BACKGROUND: Indigenous chickens could serve as precious genetic resources that should be considered in conservation and breeding programs. The Major Histocompatibility Complex (MHC) has a strong association to disease resistance/susceptibility, production and reproduction traits in chicken. Therefore, identifying its polymorphism in populations under selective breeding could be used for selection of disease resistant and higher productive breeds. MHC association with quantitative traits could be  a result of its linkage with causative genes controlling these traits. Insulin-like growth factor 1 (IGF1) is a candidate marker for phenotypic traits in chicken which are associated with important production and reproduction features. Objectives: Based on this hypothesis, MHC polymorphism and its association to IGF1 gene (as a marker for production traits) were investigated in Khorasan indigenous chicken. Methods: In total, 313 DNA samples that belonged to the Khorasan indigenous chicken were analyzed. LEI0258 microsatellite marker and fragment analysis method was used for MHC genotyping. Single nucleotide polymorphism (SNP) of the IGF1 5’-UTR was detected by restriction fragment length polymorphism (PCR-RFLP) and PstI restriction endonuclease enzyme. Linkage disequilibrium between MHC and IGF1 loci were also determined using SAS/Genetics software and likelihood ratio test. Results: Collectively, 25 different alleles (185-493 bp) and 76 genotypes of LEI0258 microsatellite were identified in Khorasan population. Two alleles, A (PstI -) and B (PstI +) and three genotypes (AA, AB and BB) were identified for IGF1 gene. Significant linkage disequilibrium (p=0.0083) was observed between LEI0258 and IGF1 loci in this population. Conclusions: These results indicate a high MHC genetic diversity in Khorasan indigenous chicken as a valuable genetic resource. Results from MHC/IGF1 linkage study confirm the hypothesis that MHC association with production traits could be as a result of MHC linkage with causative genes controlling the traits.

BACKGROUND: 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. Objectives: 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. Methods: 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. Results: 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). ConclusionS: 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.

BACKGROUND: Major histocompatibility complex (MHC) in chicken has profound influence on resistance/susceptibility to disease, and production and reproduction traits. Microsatellite marker LEI0258 is a genetic indicator for MHC haplotypes. Recognizing diversity of MHC haplotypes in selectively bred populations will be helpful for selecting population resistant to disease and development of effective vaccines. Objectives: The purpose of the present study was to evaluate polymorphism at MHC in two populations of Khorasan indigenous chickens and commercial Leghorn breed using microsatellite marker LEI0258 and to investigate its segregation and heredity. Methods: A total of 335 blood samples from Khorasan Razavi indigenous chickens and commercial Leghorn population including parents (P) and offspring (F1), were analyzed. The MHC genotypes were determined using LEI0258 microsatellite. The study of allele heredity from P to F1 and Hardy-Weinberg equilibrium were conducted using Chi-square and Likelihood Ratio tests. Results: In Khorasan indigenous chickens 20 different alleles were identified for LEI0258 microsatellite. The allele 321 bp had the highest (22.88%) and the allele 182 bp had the lowest (0.16%) frequency. In the commercial population (Leghorn breed) 3 alleles were found for this marker of which the allele 261 bp had the highest (50%) and alleles 487 bp had the lowest (6 %) frequency. In allele heredity analysis and Hardy-Weinberg equilibrium of Khorasan population, no significant differences were observed between P and F1 progenies. ConclusionS: These results indicate a higher genetic variation in indigenous chickens compared to commercial breed. There was no preference for a particular allele in indigenous chickens. The higher frequency of some alleles in F1 population is due to the high frequency of the same alleles in parent population which their gametes make the population gene pool.

Introduction: Pregnancy is a physiological state in which the immune system undergoes certain changes. On the one hand, by depleting cell defence mechanisms, it favours development and maintenance of the pregnancy. At the same time cells of the immune system ensure resistance to many risk factors, including infectious agents.Material and Methods: The study was carried out on 24 Polish Konik breed mares which were divided into two equal groups. The first group (group I) included mares living in the reserve. The second group (group II) comprised mares maintained under conventional conditions in the stables. The blood samples were collected for the first time in the perinatal period, i.e. 2 weeks before parturition (trial 0), then within the first 24 h after delivery, and then on 7ᵗʰ and 21ˢᵗ day after foaling. Flow cytometric analysis of lymphocyte expressing TCD4+, TCD8+, CD2+, and MHC class II antigens was performed.Results: Before the delivery, in group I there was a significantly higher CD4:CD8 ratio compared to group II (P ≤0.05). Similarly, significantly increased CD4:CD8 ratio in group I was noted within 24 h after parturition (P ≤0.001) and it was also observed on 7ᵗʰ day (P ≤0.03) and 21ˢᵗ day after foaling (P ≤0.02). In the first 24 h after parturition, a significant decline of lymphocytes CD8+ (P ≤0.02) was noted. No significant differences in terms of lymphocytes CD2+ and CD3+ were observed. Expression of MHC-II molecules before and after the parturition was higher in group I compared to group II; however, the difference between the groups was not significant.Conclusion: The results obtained indicate that mares living in the reserve display higher activity of cell defence mechanisms.

Based on analysis of available genome sequences, five gene lineages of MHC class I molecules (MHC I-U, -Z, -S, -L and -P) and one gene lineage of MHC class II molecules (MHC II-D) have been identified in Osteichthyes. In the latter lineage, three MHC II molecule sublineages have been identified (MHC II-A, -B and -E). As regards MHC class I molecules in Osteichthyes, it is important to take note of the fact that the lineages U and Z in MHC I genes have been identified in almost all fish species examined so far. Phylogenetic studies into MHC II molecule genes of sublineages A and B suggest that they may be descended from the genes of the sublineage named A/B that have been identified in spotted gar (Lepisosteus oculatus). The sublineage E genes of MHC II molecules, which represent the group of non-polymorphic genes with poor expression in the tissues connected with the immune system, are present in primitive fish, i.e. in paddlefish, sturgeons and spotted gar (Lepisosteus oculatus), as well as in cyprinids (Cyprinidae), Atlantic salmon (Salmo salar), and rainbow trout (Oncorhynchus mykiss). Full elucidation of the details relating to the organisation and functioning of the particular components of the major histocompatibility complex in Osteichthyes can advance the understanding of the evolution of the MHC molecule genes and the immune mechanism.

Thrombocytes in vertebrates other than mammals, inter alia in fish, are analogues of platelets in mammals. In Osteichthyes, these cells take part in haemostatic processes, including aggregation and release reactions in cases of blood vessel damage, and in the immune response development as well. This paper discusses the development of thrombocytes in Osteichthyes, taking into account the need to make changes to the concept of grouping progenitor cells as suggested in the literature. The following pages present the morphological and cytochemical properties of thrombocytes as well as their defence functions, and also point out differences between thrombocytes in fish and platelets in mammals. The paper further highlights the level of thrombocytes’ immune activity observed in fish and based on an increased proportion of these cells in response to antigenic stimulation, on morphological shifts towards forms characteristic of dendritic cells after antigenic stimulation and on the presence of surface structures and cytokines released through, inter alia, gene expression of TLR receptors, MHC class II protein-coding genes and pro-inflammatory cytokines. The study also points out the need to recognise thrombocytes in Osteichthyes as specialised immune cells conditioning non-specific immune mechanisms and playing an important role in affecting adaptive immune mechanisms.

Objective—To determine effects of infection with bovine leukosis virus (BLV) on lymphocyte proliferation and apoptosis in dairy cattle. Animals—27 adult Holstein cows. Procedures—Peripheral blood mononuclear cells (PBMCs) were isolated from whole blood from lactating Holstein cows seronegative for BLV (n = 9 cows), seropositive for BLV and aleukemic (aleukemic; 9), and seropositive for BLV and persistently lymphocytotic (PL; 9). Isolated PBMCs were assayed for mitogen-induced proliferation and were analyzed by means of flow cytometry. The PBMCs from a subset of each group were assayed for apoptosis, caspase-9 activity, and expression of selected genes related to apoptosis. Results—PL cows had significantly higher total lymphocyte counts and significantly lower proportions of T-lymphocyte populations than did BLV-negative and aleukemic cows. Both groups of BLV-infected cows had significantly higher proportions of B cells and major histocompatibility complex II–expressing cells than did BLV-negative cows. Proliferation with concanavalin A was significantly lower for PL cows, compared with proliferation for BLV-negative cows. Pokeweed mitogen–induced proliferation was significantly higher for aleukemic and PL cows than for BLV-negative cows. Gene expression of apoptosis-inhibitory proteins BCL2 and BCL2L1 was significantly higher for aleukemic cows and expression of BCL2 was significantly higher for PL cows than for BLV-negative cows. Conclusions and Clinical Relevance—Cattle infected with BLV had marked changes in PBMC populations accompanied by alterations in proliferation and apoptosis mechanisms. Because the relative distribution and function of lymphocyte populations are critical for immune competence, additional studies are needed to investigate the ability of BLV-infected cattle to respond to infectious challenge.

Total serum hemolytic complement (CH50) activity was determined for 3 semi-inbred strains of miniature swine (SLAa, SLAc, SLAd) and 1 recombinant strain SLAg (ABCcDd), each homozygous for a distinct major histocompability complex haplotype. Initial determination was made at 8 weeks of age, prior to standardized immunization, the second at age 12 weeks, after immunization. Analysis of variance was by least-squares method, using a linear model on data from 33 litters by 14 sires and 16 dams. Analysis of variance indicated that the combined effects of haplotype, sire, dam, litter, and gender accounted for 47.63% of the total variation in preimmunization CH50 values. Dam (P less than or equal to 0.06) and litter (P less than or equal to 0.03) significantly influenced preimmunization complement activity. Although swine leukocyte antigen (SLA) haplotype was not significant in the model, least-squares mean comparisons between haplotypes suggested that ac, dg, and gg pigs tended to have comparatively low preimmunization CH50 values. The model did not account for significant variability in postimmunization CH50 values, but least-squares means indicated that dd, dg, and gg haplotypes tended to have lower values than did other haplotypes tested. Mean CH50 units for 8-and 12-week-old pigs were 41.32 +/- 20.49 and 59.50 +/- 54.35, respectively. There was a significant difference (P less than or equal to 0.001) in CH50 activity between 8- and 12-week-old pigs associated with immunization, because CH50 of nonimmunized controls did not differ at 8 and 12 weeks.

OBJECTIVE To assess efficiency of gravity filtration to enhance recovery of equine bone marrow elements including stem and progenitor cells. ANIMALS 12 healthy adult horses. PROCEDURES Bone marrow aspirates were collected from the fifth sternebral body and filtered by gravitational flow to obtain bone marrow elements. Raw and harvested bone marrow and marrow effluent were evaluated for WBC and platelet counts, automated and cytomorphologic cell differential counts, mesenchymal stem cell CFUs, cell viability, and differentiation capacity. Isolated cells were analyzed for CD90 and major histocompatibility complex (MHC) class I and II antigens. RESULTS Mean cell viability of harvested bone marrow was 95.9%. Total WBCs and platelets were efficiently captured on the filter (> 95%), and mean recovery in harvested bone marrow was 30%. Cytologic cell differential counts indicated that the percentage of neutrophils was significantly less and the progenitor cell population was significantly higher and concentrated 1.56-fold in harvested bone marrow, compared with results for raw bone marrow. Flow cytometry and cell culture were used to characterize harvested bone marrow cells as positive for expression of CD90 and negative for MHCI and MHCII, which indicated stem cells with a multipotent phenotype that differentiated into chondrocytes, osteocytes, adipocytes, and tenocytes. CONCLUSIONS AND CLINICAL RELEVANCE Gravitational filtration of bone marrow efficiently yielded platelets and cells and produced a progenitor-enriched, leukocyte-reduced product, compared with raw bone marrow.

Objective-To evaluate a model for atopic dermatitis (AD) and to measure the effect of sensitization in Beagles genetically predisposed to produce high serum concentrations of allergen specific IgE. Animals-22 laboratory Beagles. Procedure-Seventeen dogs were sensitized from birth to 3 allergens (recombinant birch pollen, Dermatophagoides pteronyssinus, and D farinae). Five nonsensitized dogs from the same litters served as controls. Clinical scoring, regular intradermal testing, measurement of serum concentrations of allergen-specific IgE, and collection of biopsy specimens of skin at 23, 32, and 43 weeks of age were performed. Serial tissue sections were stained for identification of IgE+ cells, mast cells and their subtypes, T-cells, Langerhans cells, and major histocompatibility complex class-II+ cells. At the age of 15 months, dogs were continuously exposed to 2 µg of mite allergen/ g of dust. Results-Sensitized dogs had positive intradermal test reactions and significantly higher serum concentrations of allergen specific IgE, compared with nonsensitized dogs. In sensitized and nonsensitized dogs, a significantly higher number of mast cells was found at predilection sites, compared with the control biopsy site. The number of mast cells at predilection sites increased with age. Sensitization significantly increased the number of epidermal Langerhans cells by 23 weeks of age. The number of epidermal Langerhans cells significantly increased in nonsensitized dogs by 32 weeks of age. Clinical scoring only revealed mild transient erythema in some dogs. Conclusion and Clinical Relevance-Increases in concentrations of serum allergen-specific IgE and exposure to allergens is not sufficient to induce clinical signs of AD in genetically predisposed dogs.