gholamraza nikbakhat brujeni

Major histocompatibility complex (MHC) encodes for highly variable molecules, most of which are responsible for foreign antigen recognition and activation of immune responses in the host. LEI0258 microsatellite, located in the poultry MHC region, is a suitable genetic marker for determining MHC haplotypes and genetic diversity in poultry.

Considering the fact that there is no report on the frequency and types of MHC alleles and population genetic analysis in Ross 308 poultry in Iran, the present study aimed to investigate the diversity of MHC haplotypes of Ross 308 broilers by LEI0258 microsatellite.

A total of 216 blood samples were collected from two productive herds of Ross 308 broilers. After extracting DNA of the blood samples and amplifying LEI0258 microsatellite alleles, genotyping of MHC haplotypes was performed using agarose gel electrophoresis and fragment analysis techniques.

A total of seven alleles and 21 genotypes were identified for LEI0258 microsatellite in these two groups. the highest and the lowest frequencies belonged respectively to allele 385 bp (42.86 %) and allele 300 bp (4.33 %). Heterozygous 207/385 was found to be the dominant genotype in both populations. According to the similarity matrix analysis, there was an 84.56 % similarity between the two groups.

The results obtained in this study revealed a high level of heterozygosity (85.71 % and 91.35 %) and deviation from Hardy–Weinberg equilibrium (P<0.0001) in these two Ross populations. Ross 308 broiler chickens had lower allelic diversity and higher genetic similarity compared to the native ones. These findings provided additional information on the use of MHC as a candidate gene marker in genetic improvement and resource conservation in broiler populations.

Magnetic cell targeting is a novel non-invasive cellular delivery technique. It improves stem cell delivery to and retention in the injury site. Labeling cells with superparamagnetic iron oxide nanoparticle (SPION) is one of the most important steps of this technique. Appropriate SPIONs selection is believed to be of vital importance.

The current study aimed to produce SPIONs which are capable of attaching to Mesenchymal stem cells surface (MSCs).

Dextran coated SPIONs were produced following co-precipitation method under N2 atmosphere. Bone marrow derived MSCs were isolated and cultured from rabbit humerus bone. Anti-rabbit CD44 monoclonal antibody was attached to the surface of SPIONs and MSCs and were labeled with this final product. SPIONs coating process, particle size, and antibody conjugation efficacy were evaluated using FT-IR, SEM, and Bradford protein measurement assay, respectively. Attachment of antibody-linked dextran coated SPIONs to MSCs was accessed utilizing Prussian blue staining, immunofluorescence analysis, and SEM analysis.

Peaks of FT-IR at 3200 cm-1 and 2922 cm-1 are representative of dextran. The average particle size was 56.13±6.67. The average antibody-SPION conjugation ratio was 77.78±6.35%. The average percentage of the labeled cells in Prussian blue and IF analysis were 71.57±2.53 and 95.04±0.95, respectively. MSCs-SPIONs conjugation was also confirmed via SEM analysis.

In conclusion, it could be inferred that mesenchymal stem cells could successfully be labeled with dextran coated-anti CD44 antibody conjugated- superparamagnetic Iron oxide nanoparticles. This product could be used for further in-vitro and in-vivo evaluations.

Extensive effort is focused on identifying genomic conserved antigens in development of effective vaccine against Enterotoxigenic Escherichia coli. Antigen 43 is one of the members of a large secreted protein family named autotransporters in the E.coli and other gram negative bacteria. Autotransporter proteins have a similar conserved structure. Some of them are recognized during both experimental and naturally occurring ETEC infections. Antigen 43 is represented as a potential target in vaccine development because of its virulence functions such as cell aggregation, biofilm formation and its presence in convalescent sera from human patients with ETEC diarrhea. In this study, we carefully investigate antigenic structure and immunogenicity of the Antigen 43 protein of strain 510 E.coli isolated from calves by experimental methods and immunoinformatics tools. Amino acid sequence, physico-chemical parameters, stability, secondary and tertiary protein structure, the ability of induction the B and T cell immune responses by having the effective epitopes and also the allergenicity assessment were analyzed. In this study, we identified 15 peptide sequences that can potentially induce B and T cell immune responses and finally, 9 of them were introduced as antigens. The results of in-silico analysis on this protein suggested that it can be used in bovine colibacillosis vaccine development.

Major histocompatibility complex (MHC) is one of the best characterized genetic regions controlling immune responses against vaccines. Identifying the association between MHC haplotypes and improved immune responses would be useful in genetic breeding strategies in animals. MHC class II B genetic diversity and its association with humoral immune responses against Newcastle vaccine (NDV) were evaluated in commercial turkey poults (meleagris gallopavo). A total of 92 turkey poults were vaccinated with live VG/GA strain of Newcastle disease vaccine at age of 10 and 20 days. Serum NDV specific IgY was assessed by indirect enzyme linked immunosorbent assay (ELISA) and MHC II B polymorphism was determined using high resolution melting curve (HRM) technique and DNA sequencing method. Effects of alleles on humoral immune responses were evaluated by multivariate regression analysis and GLM procedures. A total of 8 HRM profiles and 10 alleles were identified in this population. B1-1*3, B1-1*4 and B1-1*5 alleles were significantly associated with lower antibody responses against vaccine in commercial turkey poults. Three alleles reported in this study were associated with reduced immune responses against NDV vaccine in turkey population. Due to such negative associations, molecular breeding programs based on specific genetic markers should be implemented with great caution.

Diarrhea is a common disease in the neonate calf which imposes significant economic burden on cattle industry around the world. During the first week after birth, Enterotoxigenic E.coli (ETEC) strains carrying F5 fimbria are one of the most important pathogens causing calf diarrhea. F5 fimbria is involved in early stage of pathogenesis and is responsible for attachment of bacteria to enterocytes; this attachment is mediated by FanC protein of F5 fimbria. Antibodies directed against F5 fimbriae play a significant role in prevention and control of the disease. Evaluation and expression of recombinant expression of F5 Fimbriae of Enterotoxigenic Escherichia Coli associated with calf diarrhea. In the present study, the fanC region of F5 fimbria was cloned in a pET28a plasmid. The recombinant construct was confirmed by sequencing and protein production in Escherichia coli BL21 (DE3) was evaluated by western blotting procedure. Based on our findings, the recombinant FanC protein or the BL21 (DE3) strain are suitable candidates to develop an effective vaccine against calf colibacillosis or use in a diagnostic kit for F5+ ETEC.

The function of molecules of major histocompatibility complex (MHC) is to bind antigenic peptides and display them on the cell surface for recognition by T cells. MHC polymorphism is related to suseptibility or resistance to immune-mediated and infectious diseases. High Resolution Melt (HRM) analysis is a novel and powerful technique in molecular biology for the detection of genetic polymorphisms and can be used for discrimination of MHC alleles and genotyping.
Discrimination of dog MHC alleles (DLA-DRB1) by high-resolution melt analysis.
Forty blood samples were collected from dogs referred to Small Animal Teaching Hospital of faculty of veterinary medicine, University of Tehran. After DNA extraction and amplification the exon 2 of DLA-DRB1, HRM and Melt Curve analysis was carried out. Direct sequencing was used for checking the HRM patterns.
Following the HRM and Melt curve analysis, forty samples could be classified into 8 genotypes (A to H). Most frequent was A (25.00%) and then C and E (each one 15.00%). In total, 82.5% and 17.5% of samples were heterozygote and homozygote, respectively.
In this study, the HRM technique was used for DLA-DRB1 gene alleles typing for the first time. Confirmation HRM results through sequencing results revealed that HRM can be used for discrimination of DLA-DRB1 alleles.

Major histocompatibility complex (MHC) comprises a group of genes, which plays a central role in immune response. The exon 2 of BuLA-DRB3 is part of the MHC class II in buffalo that highly polymorphic, found to be associated with resistance/susceptibility to infections and also with production parameters.
The purpose of the present study is to identify BuLA-DRB3 polymorphism in Khuzestan buffaloes and compare this population with other Iranian and world buffalo populations.
Blood samples were taken from 136 unrelated Khuzestan river buffaloes. After DNA extraction, second exon of BuLA-DRB3 was amplified by the seminested PCR method. Then, the fragments produced by amplifying second exon were cut by RsaI restriction enzyme according to van Eijik method. In the following, allelic frequencies, genotype frequencies, expected and observed homozygosty and heterozygosity were calculated.
In restriction fragment analysis 13 and 24 different allelic and genotype patterns were identified for RsaI restriction enzyme, respectively. 10 out of 13 alleles were previously reported. The most frequent genotype was oo)0.1691) and then followed by hh (0.1544) ,ll(0.1103), lw (0.0955), lh (0,0808), ha (0.0661) and lo (0.0514). Also four most frequent alleles were o (0.2721), h (0.2316),l (0.2316)and w (0.1176), respectively. These seven genotypes and four alleles form 72.76% and74.29% overall genotype and allele frequency of population. In addition, estimation of heterozygosity/homozygosty and deviation from Hardy-Weinberg equilibriumofcorresponding population revealed observed homozygosty is more than heterozygosity and departure of population from Hardy-Weinberg equilibrium.
The results indicated that exon 2 of the BuLA-DRB3 gene is highly polymorphic among Khuzestan buffaloes and although, there is differences between buffalo’s genetic polymorphism of distinct world regions, Khuzestan buffaloes’ population is similar to Egyptian buffaloes’ population.

Major histocompatibility complex (MHC) plays a central role in regulation and control of the immune responses to infectious diseases. Due to its polymorphism, individual differences in response to vaccines have been observed in different chicken populations. Studying the association of chicken MHC with immune response to vaccines will help the control of infectious disease and vaccination success. The present study aimed to evaluate the MHC polymorphism and its association with antibody response against infectious bursal disease (Gumboro), Newcastle (ND) and Influenza (AI) vaccines in Khorasan native chickens. Diversity of LEI0258 microsatellite marker (MHC genotyping) was investigated by fragment analysis method. Antibody titer against IBD was measured by ELISA and antibody titers against ND and AI vaccines were measured by Haemaglutination Inhibition (HI) assay. Statistical analysis was performed using SPSS software (version 21). Univariate regression analysis was performed using weighted least squares with weight number of progeny mean data. Total of 13 LEI0258 microsatellite alleles were identified in Khorasan native chickens which indicated a high genetic diversity in the population. The allele 361 bp had the highest (28.48%) and the allele 350 bp had the lowest (0.69%) frequency, respectively. In evaluating the association of MHC with immune responses, 311 and 313 bp alleles were significantly associated with elevated immune responses to Newcastle vaccine, while allele 266 bp was associated with lower IBDV antibody titers (p<0.05). According to the important role of MHC in controlling infectious disease resistance or susceptibility and quality of immune responses, these results could be used for selection and improving the populations under selective breeding.