Publications by authors named "Ahmad Mahmoudi"

27 Publications

  • Page 1 of 1

The complete mitochondrial genomes of three mole vole species (Rodentia: Arvicolinae).

Mitochondrial DNA B Resour 2020 Jun 17;5(3):2485-2487. Epub 2020 Jun 17.

Department of Molecular Systematics, Zoological Institute RAS, St. Petersburg, Russia.

The subterranean voles of the genus are species of subfamily Arvicolinae well adapted to underground life. In this paper, we report the assemblies of complete mitochondrial genomes for three mole voles from genus - northern mole vole (16,376 bp), transcaucasian mole vole (16,540 bp), and southern mole vole s (16,388 bp). Each of three mitogenomes encode for 12S and 16S rRNAs, 22 tRNAs, 13 protein-coding genes, and D-loop in the characteristic arrangement of subfamily Arvicolinae (Rodentia: Cricetidae). This study verifies the evolutionary status of subgenera and within the genus at the molecular level. The mitochondrial genome would be a significant supplement for the genetic background. The three species formed a monophyletic group with the high bootstrap value (100%) in all examinations.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1080/23802359.2020.1778567DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7782971PMC
June 2020

Plague reservoir species throughout the world.

Integr Zool 2020 Dec 2. Epub 2020 Dec 2.

Department of Epidemiology and Biostatistics, Research Centre for Emerging and Reemerging Infectious Diseases, Pasteur Institute of Iran, Tehran, Iran.

Plague has been known since ancient times as a re-emerging infectious disease, causing considerable socioeconomic burden in regional hotspots. To better understand the epidemiological cycle of the causative agent of the plague, its potential occurrence, and possible future dispersion, one must carefully consider the taxonomy, distribution, and ecological requirements of reservoir-species in relation either to natural or human-driven changes (e.g. climate change or urbanization). In recent years, the depth of knowledge on species taxonomy and species composition in different landscapes has undergone a dramatic expansion, driven by modern taxonomic methods such as synthetic surveys that take into consideration morphology, genetics, and the ecological setting of captured animals to establish their species identities. Here, we consider the recent taxonomic changes of the rodent species in known plague reservoirs and detail their distribution across the world, with a particular focus on those rodents considered to be keystone host species. A complete checklist of all known plague-infectable vertebrates living in plague foci is provided as a Supporting Information table.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1111/1749-4877.12511DOI Listing
December 2020

Infection of hard ticks in the Caspian Sea littoral of Iran with Lyme borreliosis and relapsing fever borreliae.

Ticks Tick Borne Dis 2020 11 1;11(6):101500. Epub 2020 Jul 1.

Department of Epidemiology and Biostatistics, Research Centre for Emerging and Reemerging Infectious Diseases, Pasteur Institute of Iran, Tehran, Iran; National Reference Laboratory for Plague, Tularemia and Q Fever, Research Centre for Emerging and Reemerging Infectious Diseases, Pasteur Institute of Iran, Akanlu, Kabudar Ahang, Hamadan, Iran.

The Caspian Sea littoral of Iran is home to various hard tick species, including Ixodes ricinus, the notorious vector of Lyme borreliosis (LB) in Eurasia. Here, in this area, we examined I. ricinus and other hard ticks, along with common rodents and small mammals for LB and relapsing fever (RF) borreliae infection. Ticks were collected from various mammalian hosts, including sheep, goats, cattle, camels, horses, dogs, donkeys, rodents, and hedgehogs. Rodents and small mammals were live-captured from different habitats. A real-time PCR for 16S rRNA sequence revealed borrelial DNA in 71 out of 501 (≈14 %) specimens belonging to I. ricinus and Rhipicephalus ticks. None of the rodents and small mammals showed borrelial infection in the viscera. PCR amplification and sequencing of a 600-bp sequence of the flaB identified Borrelia bavariensis, Borrelia garinii, Borrelia afzelii, and Borrelia valaisiana, and the RF Borrelia, B. miyamotoi in I. ricinus ticks. The RF-like Borrelia in Rhipicephalus ticks shared the highest identity (98.97 %) with an isolate infecting Haemaphysalis megaspinosa ticks in Japan. Our phylogeny and BLAST analysis suggest the range extension of the European I. ricinus-associated borreliae into the north of Iran.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1016/j.ttbdis.2020.101500DOI Listing
November 2020

Human immune response to salivary gland antigens in a leishmaniasis-endemic focus in Iran.

Pathog Glob Health 2020 09 9;114(6):323-332. Epub 2020 Jul 9.

Department of Medical Entomology and Vector Control, School of Public Health, Tehran University of Medical Sciences , Tehran, Iran.

Salivary proteins specific antibodies have been shown to be useful biomarkers of exposure to sand fly bites. This study aimed to investigate the level, duration, and dynamics of the human immune response against the SGL of  Parrot, 1917 (Diptera: Psychodidae), and to assess the immunoreactivity of human sera with SGL components in an endemic area of anthroponotic cutaneous leishmaniasis (ACL) in Iran. The study was carried out in 2-phase; longitudinal and cross-sectional. Sand flies were collected monthly from indoors and outdoors. In the longitudinal study, sera from healthy volunteers were collected monthly, and in the cross-sectional study, sera from healthy volunteers and patients with ACL lesion/s, were collected for immunoassay studies. The level of anti- saliva IgG was detected using the ELISA. Immunoreactivity of individual human sera with saliva components was also assessed by western blotting. was the predominant sand fly species in the study area. The maximum and minimum percentages of IgG responses were seen in October (66%) and March (29%), respectively. Additionally, the cross-sectional study showed that 59.3% of the healthy volunteers and 80% of the patients were IgG positive. The antibody response against  salivary gland was high during the sand fly active season and declined by the end of the activity of the vectors.  Antibody response against the SGL components of  was transient and individual-specific. Some individuals shared a strong reaction against certain individual antigens, which could be considered as vector exposure markers for further investigation.

List Of Abbreviations: ELISA: Enzyme-Linked Immunosorbent Assay; SDS PAGE: Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis; SGL: Salivary Gland Lysate; ACL: Anthroponotic Cutaneous Leishmaniasis; PBS: Phosphate Buffered Saline; BCA: Bicinchoninic Acid; PBS-T: Phosphate Buffered Saline Tween; FBS: Fetal Bovine Serum; HRP: Horseradish Peroxidase; TMB: 3,3',5,5'-Tetramethylbenzidine; PVDF: Polyvinylidene Difluoride; SGA: Salivary Gland Antigens; OD: Optical Density; KDa: Kilodalton; VL: Visceral Leishmaniasis; CL: Cutaneous Leishmaniasis; SGs: Salivary glands.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1080/20477724.2020.1789399DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7480592PMC
September 2020

Wild Rodents and Their Ectoparasites in an Enzootic Plague Focus, Western Iran.

Vector Borne Zoonotic Dis 2020 05 20;20(5):334-347. Epub 2020 Feb 20.

National Reference Laboratory for Plague, Tularemia and Q Fever, Research Centre for Emerging and Reemerging Infectious Diseases, Pasteur Institute of Iran, Akanlu, Kabudar Ahang, Hamadan, Iran.

Entomological surveys of ectoparasites and their hosts are an essential tool for assessing the risks of rodent-borne diseases transmitted to humans by arthropod vectors. This study was carried out to update the epidemiological data of plague with respect to species compositions of the rodents and their ectoparasites at enzootic foci located in Kurdistan Province, Iran. The rodents' habitats were selected based on past records of plague and subclimates in each study district with especial attention to the vegetation type. The trapped rodents were anesthetized using a chloroform chamber, and the animals were then examined for ectoparasites by brushing their hair over a pan containing water. The ectoparasites were collected with a fine brush and preserved in 70% ethanol in screw cap tubes. A total of 208 rodents were trapped from three districts. Taxonomic ranking of the rodents indicated that the specimens belonged to 2 suborders of Myomorpha and Sciuromorpha, 4 families (Muridae, Muscardinidae, Cricetidae, and Sciuridae), 7 genera, including , , , , , and , and 15 species. Out of 208 rodents, only 56 (26.9%) were infested with 22 species of ectoparasites. Totally, 312 ectoparasites were isolated from 56 rodents, including 12 flea species (54.5%), 6 mite species (27.3%), 3 tick species (13.6%), and one louse species (4.6%). Five species of fleas were recorded for the first time in Kurdistan Province, including , , , sp., and spp. The finding revealed new records for rodents and ectoparasites in Kurdistan Province, as well as changes in dominant rodent species and their ectoparasites compared to previous studies. This phenomenon can influence the changes in the incidence of plague and its epidemiology.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1089/vbz.2019.2524DOI Listing
May 2020

A serological and molecular study on Francisella tularensis in rodents from Hamadan province, Western Iran.

Comp Immunol Microbiol Infect Dis 2020 Feb 5;68:101379. Epub 2019 Nov 5.

National Reference Laboratory for Plague, Tularemia and Q Fever, Research Centre for Emerging and Reemerging Infectious Diseases, Pasteur Institute of Iran, Kabudar Ahang, Hamadan, Iran; Department of Epidemiology and Biostatistics, Research Centre for Emerging and Reemerging Infectious Diseases, Pasteur Institute of Iran, Tehran, Iran. Electronic address:

Introduction And Purpose: Tularemia is a zoonotic disease, the most important hosts of which are rodents. Endemic regions and reservoirs of F. tularensis are not well-researched areas in Iran. The present study aimed to study F. tularensis infection in the rodent populations of western Iran.

Materials And Methods: Samples were collected in different areas of Kabudar Ahang County in Hamadan province (west of Iran) from 2014 to 2017. Tularemia serological and molecular tests were conducted using the tube agglutination test and Real-time PCR method tracking the ISFtu2 gene. Positive serum samples were evaluated for cross-reactivity with brucellosis.

Results: A total of 433 rodents, collected from 33 localities, were included in the study. The most abundant species belonged to the Persian jird (Meriones persicus; 75.5%), and Libyan jird (Meriones libycus; 10.1%). Among the studied samples, three (0.74 %) were seropositive and five (1.15%) were PCR positive. Seropositive samples were two M. persicus and one M. libycus, and PCR positive rodents were four M. persicus and one M. vinogradovi. Tularemia seropositive samples showed no cross-reactivity with brucellosis.

Conclusion: Given the presence of infection in rodents with tularemia agent in the studied area, it is crucial to elucidate the risks of rodent exposure to tularemia for physicians, health personnel and the general population.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1016/j.cimid.2019.101379DOI Listing
February 2020

Hantavirus infection in Iranian patients suspected to viral hemorrhagic fever.

J Med Virol 2019 10 5;91(10):1737-1742. Epub 2019 Jul 5.

Department of Epidemiology and Biostatistics, Research Centre for Emerging and Reemerging Infectious Diseases, Pasteur Institute of Iran, Tehran, Iran.

Background: Hantaviruses are a group of emerging pathogens causing hemorrhagic fever with renal syndrome and Hantavirus cardiopulmonary syndrome in human. This study was conducted to investigate Hantavirus infection among Iranian viral hemorrhagic fever suspected patients.

Methods: From April 2014 to June 2016, 113 cases from 25 different provinces of Iran were analyzed for Hantavirus infection by IgM/IgG ELISA and pan-Hantavirus RT-PCR tests.

Results: Although, viral genome was detected in none of the subjects, IgM and IgG antibodies were detected in 19 and 4 cases, respectively. Differentiation of the anti-Hantavirus antibodies according to virus species by EUROLINE Anti-Hantavirus Profile Kit revealed three Puumala virus IgM positive, one Hantaan virus IgM positive, one Hantaan virus IgM borderline, and two Puumala virus IgG borderline cases.

Conclusions: This study demonstrates the circulation of Hantaviruses in Iran and calls for further investigations of these life-threatening viruses in the country.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1002/jmv.25522DOI Listing
October 2019

Rodents Helminth Parasites in Different Region of Iran.

Iran J Parasitol 2018 Apr-Jun;13(2):275-284

Dept. of Biology, Faculty of Science, Ferdowsi University of Mashhad, Mashhad, Iran.

Background: Climate condition is expected to have significant in rodents' diversity and in the seasonal pattern of diseases carried by different rodents. In an effort to aid in the study of the biodiversity of parasites of rodents in different climate zoon we examined climate patterns in the parasite assemblages of different rodents from Mar 2015 to Feb 2016.

Methods: Of 253 captured rodents in three climate zone of Iran, thirteen species of rodents were recognized. Rodents included , , , , , and . Trapped rodents humanely sacrificed and the gastrointestinal and respiratory tracts were removed and examined to identify parasitic helminths. Parasites were identified using key morphological characteristics.

Results: Of 253 rodents examined, 109 (43.08%) were positive for helminth infection including (20.1%), (9.9%), (0.3%), sp. (0.3%), sp. (0.7%), sp. (1.1%), (6.7%) (4.3%) (3.1%), (0.8%), (2.7%), sp. larva (0.3%) and (0.3%). was the only species of Trematoda isolated from water vole () for the first time in Iran.

Conclusion: Some rodents are omnivorous, showing high predisposition to helminths parasites consequently, they harbor some species of parasites which are potentially zoonotic or may serve as vectors of important zoonotic pathogens. Therefore, the potential health hazard of these species needs to be considered to prevent infectivity of humans.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6068363PMC
August 2018

Molecular Survey of Tularemia and Plague in Small Mammals From Iran.

Front Cell Infect Microbiol 2018 10;8:215. Epub 2018 Jul 10.

Department of Clinical Microbiology and the Laboratory for Molecular Infection Medicine Sweden, Umeå University, Umeå, Sweden.

Plague and tularemia are zoonoses and their causative bacteria are circulating in certain regions of Iran. This study was conducted to investigate potential disease reservoirs amongst small wildlife species in different regions of Iran. Rodents, insectivores and hares from 17 different provinces of the country were collected in 2014 and 2015. Samples were taken from the spleens of the animals and Real-time PCR was applied to detect nucleic acid sequences that are specific to and , respectively. Among 140 collected rodents, 25 distinct species were identified out of which five were the most common: (21% out of 140 rodents), (12%), (11%), (11%) and (10%). Seventeen insectivores were collected and identified as (82%) and (18%). Fifty-one hares were collected and identified as (57%), (14%) and sp. (29%). Three out of 140 explored rodents (1.91%) were positive for , an , a , and a collected from Golestan, Khuzestan and Razavi Khorasan provinces, respectively. Two hares (3.92%) were -positive, a from Khuzestan and a sp. from the Sistan and Baluchistan province. None of the tested animals were positive for . This is the first report of direct detection of in mammals of Iran and the first-time observation of the agent in a snow vole, worldwide. The results indicate that tularemia is more widespread in Iran than previously reported including the Northeast and Southwestern parts of the country. Future studies should address genetic characterization of positive DNA samples from Iran to achieve molecular subtyping and rule out assay cross-reactivity with near neighbor species.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.3389/fcimb.2018.00215DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6048195PMC
July 2019

Rodent-borne diseases and their public health importance in Iran.

PLoS Negl Trop Dis 2018 04 19;12(4):e0006256. Epub 2018 Apr 19.

Department of Epidemiology and Biostatistics, Research Centre for Emerging and Reemerging Infectious Diseases, Pasteur Institute of Iran, Tehran, Iran.

Background: Rodents are reservoirs and hosts for several zoonotic diseases such as plague, leptospirosis, and leishmaniasis. Rapid development of industry and agriculture, as well as climate change throughout the globe, has led to change or increase in occurrence of rodent-borne diseases. Considering the distribution of rodents throughout Iran, the aim of this review is to assess the risk of rodent-borne diseases in Iran.

Methodology/principal Finding: We searched Google Scholar, PubMed, Science Direct, Scientific Information Database (SID), and Magiran databases up to September 2016 to obtain articles reporting occurrence of rodent-borne diseases in Iran and extract information from them. Out of 70 known rodent-borne diseases, 34 were reported in Iran: 17 (50%) parasitic diseases, 13 (38%) bacterial diseases, and 4 (12%) viral diseases. Twenty-one out of 34 diseases were reported from both humans and rodents. Among the diseases reported in the rodents of Iran, plague, leishmaniasis, and hymenolepiasis were the most frequent. The most infected rodents were Rattus norvegicus (16 diseases), Mus musculus (14 diseases), Rattus rattus (13 diseases), Meriones persicus (7 diseases), Apodemus spp. (5 diseases), Tatera indica (4 diseases), Meriones libycus (3 diseases), Rhombomys opimus (3 diseases), Cricetulus migratorius (3 diseases), and Nesokia indica (2 diseases).

Conclusions/significance: The results of this review indicate the importance of rodent-borne diseases in Iran. Considering notable diversity of rodents and their extensive distribution throughout the country, it is crucial to pay more attention to their role in spreading infectious diseases for better control of the diseases.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1371/journal.pntd.0006256DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5908068PMC
April 2018

A Field Study of Plague and Tularemia in Rodents, Western Iran.

Vector Borne Zoonotic Dis 2017 04 6;17(4):247-253. Epub 2017 Feb 6.

8 Yersinia Research Unit, National Reference Laboratory, WHO Collaborating Center for Yersinia , Institut Pasteur, Paris, France .

Introduction: Kurdistan Province in Iran is a historical focus for plague and tularemia. This study aimed at assessing the current status of these two foci by studying their rodent reservoirs.

Materials And Methods: Rodents were trapped and their ectoparasites were collected. The genus and species of both rodents and ectoparasites were determined. Serological analyses of rodent blood samples were done by enzyme-linked immunosorbent assay for plague and by standard tube agglutination assay for tularemia. Rodent spleen samples were subjected to bacterial culture, microscopic examination, and real-time PCR to search for active plague or tularemia infection.

Results: During this study, 245 rodents were trapped, of which the most abundant genera were Apodemus (40%), Mus (24.49%), and Meriones (12.65%). One hundred fifty-three fleas, 37 mites, and 54 ticks were collected on these rodents. The results of all direct and indirect tests were negative for plague. Serological tests were positive for tularemia in 4.8% of trapped rodents.

Discussion: This study is the first report on the presence of tularemia infection in rodents in Western Iran. Since Meriones persicus is a known reservoir for plague and tularemia, and this rodent carried plague and tularemia vectors in Marivan and Sanandaj districts, there is a real potential for the occurrence of these two diseases in this region.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1089/vbz.2016.2053DOI Listing
April 2017

A SNP in the 3'-untranslated region of AMPKγ1 may associate with serum ketone body and milk production of Holstein dairy cows.

Gene 2015 Dec 27;574(1):48-52. Epub 2015 Jul 27.

Young Researchers and Elite Club, Isfahan (Khorasgan) Branch, Islamic Azad University, Isfahan, Iran. Electronic address:

AMPK is the key switch for providing the energy balance between cellular anabolic and catabolic processes. In this study, we aimed to screen the PRKAG1 (AMPKγ1) gene in high, moderate, and low producing Holstein dairy cows. A sample of 100 pregnant dairy cows, comprising 41 high, 33 moderate, and 26 low milk yields were selected from three large dairy herds in Isfahan province of Iran. Body condition score (BCS) was estimated before parturition while beta hydroxyl butyric acid (BHBA) as a measure of ketone bodies was measured at the fifth day postpartum. In addition, using three primer pairs covering exons 2-11 and 3'-UTR of the PRKAG1 gene, a random sample of 10 high milk yield dairy cows were amplified and sequenced. The sequencing results showed the presence of a T12571C mutation in intron 6 and a T14280C mutation in the 3'-untranslated region (UTR) of the PRKAG1 gene. Following a PCR reaction for amplification of the 3'-UTR amplicons, single strand conformation polymorphism (SSCP) assay was implemented for discrimination of the mutation in the studied population. Then, we evaluated if the mutation associates with the BCS, serum BHBA level, and production traits. The experimental analysis showed that the mutated allele significantly increased the BHBA level, BCS, as well as milk and protein yield. Bioinformatic study revealed that this 3'-UTR mutation distorts the target site of mir-423-5p microRNA which is one of the most highly expressed microRNAs in the bovine mammary gland, liver, and kidney. Given the role of AMPK in energy metabolism, the newly identified 3'-UTR mutation highlights the importance of AMPK and suggests a role of miRNAs for regulation of cellular metabolism, metabolism disorders, and production traits in Holstein dairy cows.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1016/j.gene.2015.07.077DOI Listing
December 2015

Production and Characterization of Monoclonal Antibodies against Human Prostate Specific Antigen.

Avicenna J Med Biotechnol 2015 Jan-Mar;7(1):2-7

Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR, Tehran, Iran.

Background: Prostate Specific Antigen (PSA) is an important laboratory marker for diagnosis of prostatic cancer. Thus, development of diagnostic tools specific for PSA plays an important role in screening, monitoring and early diagnosis of prostate cancer. In this paper, the production and characterization of a panel of murine monoclonal antibodies (mAbs) against PSA have been presented.

Methods: Balb/c mice were immunized with PSA, which was purified from seminal plasma. Splenocytes of hyperimmunized mice were extracted and fused with Sp2/0 cells. By adding selective HAT medium, hybridoma cells were established and positive clones were selected by ELISA after four times of cloning. The isotypes of produced mAbs were determined by ELISA and then purified from ascitic fluids using Hi-Trap protein G column. The reactivities of the mAbs were examined with the purified PSA and seminal plasma by ELISA and western blot techniques. Furthermore, the reactivities of the mAbs were assessed in Prostate Cancer (PCa), Benign Prostatic Hyperplasia (BPH) and brain cancer tissues by Immunohistochemistry (IHC).

Results: Five anti-PSA mAbs (clones: 2G2-B2, 2F9-F4, 2D6-E8, IgG1/К) and clones (2C8-E9, 2G3-E2, IgG2a/К) were produced and characterized. All mAbs, except 2F9-F4 detected the expression of PSA in PCa and BPH tissues and none of them reacted with PSA in brain cancer tissue in IHC. Besides, all mAbs could detect a protein band around 33 kDa in human seminal plasma in western blot.

Conclusion: These mAbs can specifically recognize PSA and may serve as a component of PSA diagnostic kit in various biological fluids.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4388886PMC
April 2015

Production and characterization of a peptide-based monoclonal antibody against CD44 variant 6.

Monoclon Antib Immunodiagn Immunother 2015 Feb;34(1):36-43

1 Department of Immunology, Shiraz University of Medical Sciences , Shiraz, Iran .

The gene that codes for the CD44 family members consists of 20 exons, nine of which encode the standard form of the molecule. The other exons can be inserted in various combinations into the membrane proximal region of the extracellular domain of the protein, giving rise to variant isoforms (CD44v). CD44 variants, especially the CD44v6, have been reported to regulate tumor invasion, progression, and metastasis of carcinomas. Producing a high affinity monoclonal antibody against human CD44v6 provides a powerful tool to monitor and trace CD44v6 function in different biological fluids. In this study, a synthetic peptide from CD44v6 was conjugated to keyhole limpet hemocyanin (KLH) and injected into BALB/c mice. Splenocytes from the immunized mice were fused with murine SP2/0 myeloma cells followed by selection of antibody producing hybridoma cells. After screening of hybridoma colonies by ELISA, high affinity antibodies were selected and purified by affinity chromatography. Western blot, immunocytochemistry, and flow cytometry experiments were used to characterize the antibodies. Six stable hybridoma cell lines, designated as 1H1, 1H2, 2A12, 2G11, 3H3, and 3H7, were obtained. Flow cytometry and immunocytochemistry results showed that the new monoclonal antibodies recognized CD44v6 on the cell surface. This novel panel of anti-CD44v6 antibodies has the potential for investigating the role of CD44v6 in cancer pathogenesis.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1089/mab.2014.0077DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4350255PMC
February 2015

A Novel Monoclonal Antibody Against a Synthetic Peptide from β-Actin can React with its Corresponding Protein.

Protein Pept Lett 2015 ;22(5):419-24

Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR, Tehran, Iran.

Actin is one of the most widely studied structural and multifunctional housekeeping proteins in eukaryotic cells with important roles in many cell functions. Antibodies against β-actin and other housekeeping gene-encoded proteins are used as internal loading controls in Western blot analyses. The aim of this study was to produce a monoclonal antibody (mAb) against a synthetic peptide derived from N-terminal region of β-actin and to study its reactivity with different organisms. A synthetic peptide, derived from β-actin, was designed and used to produce a mAb by hybridoma technology. The produced antibody (clone 4E5- A10) was purified by an affinity chromatography column followed by characterization of purified mAb using SDS-PAGE, ELISA and Western blot. Our results showed that 4E5-A10 was an IgM and had desired purity and excellent reactivity with the immunizing peptide with an affinity constant of 2.7x10(8) M(-1)>. It could detect a band of about 45 kDa, corresponding to β-actin, in Western blot. Furthermore, it could react in a more sensitive manner and with a wider range of organisms than a known commercial anti β-actin antibody. Our data suggest that 4E5-A10 can act as a sensitive probe for detection of β-actin as an internal loading control, for a wide range of organisms, in Western blot analyses.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.2174/0929866522666141231111618DOI Listing
February 2016

Opticin, a small leucine-rich proteoglycan, is uniquely expressed and translocated to the nucleus of chronic lymphocytic leukemia cells.

Exp Hematol Oncol 2013 Aug 28;2(1):23. Epub 2013 Aug 28.

Immune and Gene Therapy Lab, CCK, Department of Oncology-Pathology, Karolinska Institute, Stockholm, Sweden.

Background: Opticin (OPTC) is a member of the small leucine-rich proteoglycan (SLRP) family and is localized particularly in certain extracellular matrices. We have previously reported the unique expression of another SLRP, fibromodulin (FMOD) in the leukemic cells of patients with chronic lymphocytic leukemia (CLL). OPTC is located in the same region as FMOD on chromosome 1 (1q32.1). Cluster up-regulation of genes may be observed in malignancies and the aim of the present study was to analyze the expression of OPTC in CLL cells.

Methods: The expression of OPTC was tested by RT-PCR and realtime qPCR in PBMC from CLL patients, other hematological malignancies and healthy controls. The presence of OPTC protein, and its subcellular localization, was investigated using fractionation methods where the obtained lysate fractions were analyzed by Western blotting. Deglycosylation experiments were performed to investigate the glycosylation status of the CLL OPTC.

Results: OPTC was expressed at the gene level in all patients with CLL (n = 90) and in 2/8 patients with mantle cell lymphoma (MCL) but not in blood mononuclear cells of healthy control donors (n = 20) or in tumor samples from nine other types of hematological malignancies. OPTC was detected by Western blot in all CLL samples analyzed (n = 30) but not in normal leukocytes (n = 10). Further analysis revealed a CLL-unique unglycosylated 37 kDa core protein that was found to be located preferentially in the cell nucleus and endoplasmic reticulum (ER) of the CLL cells.

Conclusions: A 37 kDa unglycosylated OPTC protein was detected in ER and in the nucleus of CLL cells and not in healthy control donors. The function of this OPTC core protein remains unclear but its CLL-specific expression and subcellular localization warrants further investigations in the pathobiology of CLL.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1186/2162-3619-2-23DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3766095PMC
August 2013

Production and characterization of a murine monoclonal antibody against human ferritin.

Avicenna J Med Biotechnol 2013 Oct;5(4):212-9

Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR, Tehran, Iran.

Background: Ferritin is an iron storage protein, which plays a key role in iron metabolism. Measurement of ferritin level in serum is one of the most useful indicators of iron status and also a sensitive measurement of iron deficiency. Monoclonal antibodies may be useful as a tool in various aspects of ferritin investigations. In this paper, the production of a murine monoclonal antibody (mAb) against human ferritin was reported.

Methods: Balb/c mice were immunized with purified human ferritin and splenocytes of hyper immunized mice were fused with Sp2/0 myeloma cells. After four times of cloning by limiting dilution, a positive hybridoma (clone: 2F9-C9) was selected by ELISA using human ferritin. Anti-ferritin mAb was purified from culture supernatants by affinity chromatography.

Results: Determination of the antibody affinity for ferritin by ELISA revealed a relatively high affinity (2.34×10(9) M (-1)) and the isotype was determined to be IgG2a. The anti-ferritin mAb 2F9-C9 reacted with 79.4% of Hela cells in flow cytometry. The antibody detected a band of 20 kDa in K562 cells, murine and human liver lysates, purified ferritin in Western blot and also ferritin in human serum.

Conclusion: This mAb can specifically recognize ferritin and may serve as a component of ferritin diagnostic kit if other requirements of the kit are met.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3838765PMC
October 2013

The protective effect of albumin on bevacizumab activity and stability in PLGA nanoparticles intended for retinal and choroidal neovascularization treatments.

Eur J Pharm Sci 2013 Nov 8;50(3-4):341-52. Epub 2013 Aug 8.

Department of Pharmaceutics, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran; Nanotechnology Research Centre, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran.

The rapidly growing applications of antibody-based therapeutics requires novel approaches to develop efficient drug delivery systems in which biodegradable polymeric nanoparticles are amongst the best candidates. In the present study bevacizumab loaded PLGA nanoparticles were formulated by water-in-oil-in-water emulsion method. Protein inactivation and aggregation are the major drawbacks of this technique. Therefore protective ability of various stabilizers was studied during entrapment process. Probable changes in VEGF₁₆₅ binding capability of bevacizumab was assayed by ELISA which portrays the antibody's bio-efficiency. Probable breakage of bevacizumab and its secondary and tertiary structural integrity upon entrapment were analyzed by SDS-PAGE and circular dichroism spectroscopy, respectively. In vitro and ex vivo released bevacizumab from the prepared nanoparticles was also investigated. Results revealed that the protein interfacial adsorption is the foremost destabilizing factor in the double emulsion method and incorporation of appropriate concentrations of albumin could protect bevacizumab against entrapment stress. Ex vivo release results, in rabbit vitreous, indicated the ability of prepared nanoparticles in prolonged release of the active antibody. Consequently this approach was an attempt to achieve sustained release PLGA nanoparticle formulation with the aim of protecting integrity and performance of entrapped bevacizumab.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1016/j.ejps.2013.07.014DOI Listing
November 2013

Distribution of vitamin D receptor and 1α-hydroxylase in male mouse reproductive tract.

Reprod Sci 2013 Apr 27;20(4):426-36. Epub 2012 Nov 27.

Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR, Tehran, Iran.

Vitamin D has been introduced as one of the main regulators of spermatogenesis. Here, for the first time, we evaluated the expression of vitamin D receptor (VDR) and 1α-hydroxylase in all organs of male mice reproductive tract by immunohistochemistry and Western blotting. Epithelial cells of epididymis, seminal vesicle, coagulating gland, ductus deferens, preputial gland, and prostate were the prominent cell types that concomitantly expressed VDR and 1α-hydroxylase. Nearly all cell types in testis expressed both proteins. Interestingly, VDR intensity in epididymis epithelial cells was reduced toward cauda, in which only strong staining of stereocilia was observed. Although been positive in caput epididymis, sperms lost their VDR expression in cauda region. In all organs, sperms failed to express 1α-hydroxylase. Specific bands of the VDR and 1α-hydroxylase were determined in all tissues, except testis in which novel unprecedented isoforms of 1α-hydroxylase were observed. Our findings could provide further convincing evidence of pivotal role of this hormone in male reproductive biology.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1177/1933719112459235DOI Listing
April 2013

A monoclonal antibody against leptin.

Hybridoma (Larchmt) 2012 Oct;31(5):372-7

Monoclonal Antibody Research Center, Avicenna Research Institute, Academic Center for Education, Culture, and Research, Tehran, Iran.

Leptin is an important protein that regulates energy storage and homeostasis in humans and animals. Leptin deficiency results in various abnormalities such as diabetes, obesity, and infertility. Producing a high affinity monoclonal antibody against human leptin provides an important tool to monitor and trace leptin function in different biological fluids. In this study, recombinant human leptin was conjugated to KLH and injected into mice. After immunization, mouse myeloma SP2/0 cells were fused with murine splenocytes followed by selection of antibody-producing hybridoma cells. After screening of different hybridoma colonies by ELISA, a high affinity antibody was selected and purified by affinity chromatography. The affinity constant of the antibody was measured by ELISA. Western blot, immunocytochemistry, and flow cytometry experiments were used to characterize the antibody. The anti-leptin antibody had a high affinity (around 1.13 × 10(-9) M) for its antigen. The saturation of the antibody with leptin (20 moles leptin per 1 mole antibody) in Western blot analysis proved that the antibody had specific binding to its antigen. Immunocytochemistry and flow cytometry on JEG-3 (human placental choriocarcinoma cell) cells revealed that the anti-leptin antibody recognized intracellular leptin. In conclusion, we report here the production and characterization of a murine anti-leptin antibody with high affinity for human leptin.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1089/hyb.2012.0045DOI Listing
October 2012

Comparison of the Photobleaching and Photostability Traits of Alexa Fluor 568- and Fluorescein Isothiocyanate- conjugated Antibody.

Cell J 2011 23;13(3):169-72. Epub 2011 Sep 23.

1. Department of Immunochemistry, Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR, Tehran, Iran.

Objective: Synthetic fluorescent dyes that are conjugated to antibodies are useful tools to probe molecules. Based on dye chemical structures, their photobleaching and photostability indices are quite diverse. It is generally believed that among different fluorescent dyes, Alexa Fluor family has greater photostability than traditional dyes like fluorescein isothiocyanate (FITC) and Cy5. Alexa Fluor 568 is a member of Alexa Fluor family presumed to have superior photostability and photobleahing profiles than FITC.

Materials And Methods: In this experimental study, we conjugated Alexa Fluor 568 and FITC dyes to a mouse anti-human nestin monoclonal antibody (ANM) to acquire their photobleaching profiles and photostability indices. Then, the fluorophore/antibody ratios were calculated using a spectrophotometer. The photobleaching profiles and photostability indices of conjugated antibodies were subsequently studied by immunocytochemistry (ICC). Samples were continuously illuminated and digital images acquired under a fluorescent microscope. Data were processed by ImageJ software.

Results: Alexa Fluor 568 has a brighter fluorescence and higher photostability than FITC.

Conclusion: Alexa Fluor 568 is a capable dye to use in photostaining techniques and it has a longer photostability when compared to FITC.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3584473PMC
March 2013

Investigating Association of Three Polymorphisms of Coagulation Factor XIII and Recurrent Pregnancy Loss.

Am J Reprod Immunol 2010 Sep;64(3):212-7

Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR, Tehran, Iran.

Problem: among important suspected causes of thrombophilia in women with recurrent pregnancy loss (RPL) are the polymorphisms of coagulation factor XIII (FXIII) gene. We performed a case-control study on the association between three polymorphisms of factor XIII (FXIII G103T, FXIII A614T and FXIII C1694T) and RPL in Iranian women.

Method Of Study: DNA samples from peripheral blood of 100 female patients with at least two recurrent abortions, as case group, and 100 healthy women with history of at least two successful deliveries were subjected to PCR-RFLP, and the frequencies of the polymorphisms were calculated and compared between the two groups.

Results: the prevalence of FXIII G103T polymorphism was 29% in the case group and 17% in the control group (P = 0.158). The frequencies of FXIII A614T and FXIII C1694T were 84% and 66% in the case group and 48% and 31% in the control group (P <0.001 and P < 0.001), respectively. The two latter polymorphisms are associated with RPL in Iranian women and increase the risk of RPL. A correlation was also found between FXIII A614T and FXIII C1694T polymorphisms (P < 0.001).

Conclusion: we suggest the evaluation of FXIII A614T and FXIII C1694T polymorphisms in women with RPL.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1111/j.1600-0897.2010.00838.xDOI Listing
September 2010

Production of Monoclonal Antibody against Human Nestin.

Avicenna J Med Biotechnol 2010 Apr;2(2):69-77

Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR, Tehran, Iran.

We have employed a peptide-based antibody generation protocol for producing antibody against human nestin. Using a 12-mer synthetic peptide from repetitive region of human nestin protein devoid of any N- or O-glyco-sylation sequences, we generated a mouse monoclonal antibody capable of recognizing human, mouse, bovine, and rat nestin. A wide variety of nestin proteins ranging from 140-250 kDa was detected by this antibody. This antibody is highly specific and functional in applications such as ELISA, flow cytometry, immunocytochemistry, and Western blot assays.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3558152PMC
April 2010

Conjugation of R-Phycoerythrin to a Polyclonal Antibody and F (ab')2 Fragment of a Polyclonal Antibody by Two Different Methods.

Avicenna J Med Biotechnol 2010 Apr;2(2):87-91

Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR, Tehran, Iran.

R-Phycoerythrin (R-PE), a fluorescent protein from phycobiliprotein family, is isolated from red algae. Conjugation of antibodies to R-PE facilitates multiple fluorescent staining methods. In the present study polyclonal antibodies and polyclonal F(ab')2 fragment antibodies were conjugated to R-PE by two different methods. The efficiency of the methods was evaluated using Immunocytochemistry (ICC) and Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS-PAGE). In the first conjugation method, PE was attached to SMCC linker followed by conjugation of antibody to PE-SMCC. In the second method, SH groups were added onto R-PE molecule, while the antibody was attached to SPDP linker. Then, the antibody-SPDP molecule was conjugated to R-PE. Our results showed that the two conjugation methods did not have any abrogative effects on the antibody binding activity.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3558147PMC
April 2010

Ectopic Expression of Sortilin 1 (NTR-3) in Patients with Ovarian Carcinoma.

Avicenna J Med Biotechnol 2009 Jul;1(2):125-31

Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR, Tehran, Iran ; Department of Cell and Molecular Biology, Khatam University, Tehran, Iran.

Gene expression profiling of ovarian carcinoma tissues has shown an increase of four-fold expression of SORT1 gene. Sortilin 1 (NTR-3) is a 95-100 kDa protein normally expressed in heart, brain, placenta, skeletal muscle, spinal cord, thyroid, and testis. However, its expression has never been reported in normal ovary. Here, we report expression of sortilin 1 in ovarian carcinoma tissues both at gene and protein levels. Sortilin 1 was expressed in all ovarian carcinoma patients (n=15) as well as ovarian carcinoma cell lines (n=5) regardless of their phenotypic characteristics. Non-malignant ovaries (n=6) did not express sortilin 1. The molecular basis for this ectopic expression is not yet clear. Our results showed a major cell surface expression of sortilin 1 rather than ER-Golgi compartment where it is mainly expressed. This finding may introduce sortilin 1 as a novel tumor marker for diagnosis of ovarian carcinoma and may signify its therapeutic value in targeted therapy.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3558127PMC
July 2009

Mutual helper effect in copulsing of dendritic cells with 2 antigens: a novel approach for improvement of dendritic-based vaccine efficacy against tumors and infectious diseases simultaneously.

J Immunother 2009 May;32(4):325-32

Department of Reproductive Immunology, Reproductive Biotechnology Research Center, Avicenna Research Institute, Shahid Beheshti University, Evin, Tehran, Iran.

To develop an efficient dendritic cell (DC)-based immunotherapy protocol, we examined whether simultaneous pulsing of DCs with a given antigen and a third-party antigen could enhance their antigen presentation capacity. Purified splenic DCs of Balb/c mice were pulsed separately with immunoglobulin G, ovalbumin, conalbumin, P15 peptide of Mycobacterium tuberculosis, and prostate-specific antigen or double combinations of the aforementioned antigens. In some settings, DCs pulsed with 1 antigen were mixed equally with those pulsed with another antigen. Antigen-pulsed DCs were injected into the footpad of syngeneic mice and proliferation of whole, CD4 and CD8 depleted lymph node cells was measured after restimulation with cognate antigen. Antigen-specific production of interferon-gamma (IFNgamma) was tested in culture supernatants. Frequency of responding lymph node cells was determined by IFNgamma enzyme-linked immunosorbent spot assay. Our results showed that copulsing of DCs with 2 unrelated antigens increased the capacity of DCs to induce antigen-specific T-cell proliferation against both antigens up to 16-fold. Injection of 2 populations of DCs each pulsed with a different antigen, increased proliferation of primed T cells significantly as well. Both CD4 and CD8 depleted populations showed vigorous proliferative response in copulsing system. In addition, copulsing of DCs with 2 antigens resulted in higher frequency of antigen-specific responding cells and significantly more IFNgamma production. Our results clearly showed that unrelated peptides and proteins could be used to enhance efficacy of DC-based vaccines and in this system, each antigen served to help the other one, a condition that we termed as "mutual helper effect."
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1097/CJI.0b013e31819aa31eDOI Listing
May 2009

Assessment of thyroglobulin expression in reproductive organs at different stages of mouse estrous cycle.

Avicenna J Med Biotechnol 2009 Apr;1(1):41-6

Department of Immunology, Faculty of Medicine, Iran University of Medical Sciences, Tehran, Iran.

Prevalence of abortion is higher in women with autoimmune thyroid disease. In the majority of cases, however, no abnormality of thyroid function is detected despite the high levels of antithyroid antibodies. The direct influence of such harmful autoantibodies in female reproductive organs may serve a role in pregnancy loss. In this study, expression of thyroglobulin in the reproductive tissues of cycling mice has been evaluated. Stages of estrous cycle were determined by cellular morphology and ratio of epithelial cells to leukocytes in vaginal smear of Balb/C mice. At each phase, the mice were sacrificed and their uterus, ovary and fallopian tubes were removed. Expression of thyroglobulin-specific transcript in endometrium was investigated by two sets of primers using reverse transcriptase-polymerase chain reaction (RT-PCR). In addition, expression of thyroglobulin in reproductive tissues was assessed by immunohistochemistry and dot blot analysis. The results showed that thyroglobulin mRNA is not expressed in endometrial tissue of Balb/C mice at any stage of estrous cycle. Immunohistochemical analysis also confirmed that thyroglobulin or its cross reactive-antigens are not expressed at the protein level in the female reproductive organs. The results showed that thyroglobulin was not expressed in the reproductive organs of female mice. It is plausible that antithyroglobulin antibodies could interact with newly-generated antigens during placentation and pregnancy.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3558115PMC
April 2009