Immunoreactivity Of Antisera Research Articles

Blood Groups ABO and Rh System Among Paternity and Kinship Cases of Iraqi Medical Legal Directorate

ABO and Rh are the most important blood group systems with various distributions reported for different populations. Traditional ABO blood group and Rh serology is based on the immunoreactivity of antisera with the carbohydrate A, B and H antigens on Red Blood Cells (RBC). Due to its medical importance in relation to different diseases, pursuing a line of investigation on the ABO and Rh blood group systems has been of significance for years. It is well known that these blood group systems are of great importance in blood transfusion and organ transplantation. Furthermore, the susceptibility to several diseases has been associated with the ABO phenotype, but such correlation remains controversial. Progress in the molecular biology of the ABO system has recognized the molecular basis of the red blood cell antigens and has provided a genetic model for ABO polymorphism at the molecular level. The ABH antigens are not primary gene product, but they are the enzymatic reaction product of enzymes called glycosyltransferases. After the ABO system, the Rh (Rhesus) blood group system is regarded as the second most important blood group system, as some of the severe hemolytic transfusion reactions and most hemolytic disease of the fetus and new born (HDFN) cases are associated with antibodies to the Rh group antigens. Objectives: To determine ABO blood group and Rh factor phenotype antigens frequency, distribution, and document among paternity cases of Iraqi Medical Legal Directorate / Clinical Forensic Department / paternity and kinship division. Materials and Methods: The material and methods were used ABO and Rh kits for detection phenotype antigens of this blood types, the principle of test is one step immunological reaction. Total samples (1997) were referred to medical Legal Directorate (MLD). Out of the total participants there were 1815 (98.4%) of samples complete information for inclusion criteria. the sample were used for estimation of phenotypic distribution of ABO, Rh. ABO blood group serology was performed at MLD, Paternity and Kinship Division using commercially available monoclonal Anti-A, Anti-B, reagents according to the national blood bank standard operating procedures. The Method principle is based on antigen antibody reaction (agglutination) some tests were confirmed under microscope examination to see weak reaction. Results: The most common blood group was O (52.3%) and the least common was AB (3.18%).97.7% of the blood participants were Rh Positive and the rest were Rh Negative. Most participants were young adults, representing the age group of 19 – 29. Most participants were male (88.1%). Conclusion: Blood group O and Rh+ factor was dominant in both sexes, Up-to-date knowledge of the distribution of blood types in a local setting is critical to the functioning of any national health service. To date, there has been a lack of data in Iraqi population.

Effect of prorenin peptide vaccine on the early phase of diabetic retinopathy in a murine model of type 2 diabetes.

Prorenin is viewed as an ideal target molecule in the prevention of diabetic retinopathy. However, no drugs are available for inhibiting activation of prorenin. Here, we tested the effect of a prorenin peptide vaccine (VP) in the retina of a murine model of type 2 diabetes (T2D). To choose the optimal vaccine, we selected three different epitopes of the prorenin prosegment (E1, E2, and E3) and conjugated them to keyhole limpet hemocyanin (KLH). We injected C57BL/6J mice twice with KLH only (as a control vaccine), E1 conjugated with KLH (E1-KLH), E2-KLH, or E3-KLH and compared antibody titers. E2-KLH showed the highest antibody titer and specific immunoreactivity of anti-sera against prorenin, so we used E2-KLH as VP. Then, we administered injections to the non-diabetic db/m and diabetic db/db mice, as follows: db/m + KLH, db/db + KLH, and db/db + VP. Retinal blood flow measurement with laser speckle flowgraphy showed that the impaired retinal circulation response to both flicker light and systemic hyperoxia in db/db mice improved with VP. Furthermore, the prolonged implicit time of b-wave and oscillatory potentials in electroretinography was prevented, and immunohistochemical analysis showed reduced microglial activation, gliosis, and vascular leakage. The enzyme-linked immunosorbent spot assay confirmed vaccinated mice had no auto-immune response against prorenin itself. The present data suggest that vaccination against prorenin is an effective and safe measure against the early pathological changes of diabetic retinopathy in T2D.

Immunogenicity Study of Plasmid DNA Encoding Mouse Cysteine‐Rich Secretory Protein‐1 (mCRISP1) as a Contraceptive Vaccine

To examine the immunocontraceptive properties of the plasmid pcDNA-mCRISP1 and compare them to the corresponding recombinant mCRISP1 (r-mCRISP1). RT-PCR and indirect immunofluorescence were performed to observe the mCRISP1 protein expression in COS-7 cells. Three groups of mice received three injections of r-mCRISP1, pcDNA-mCRISP1 or pcDNA vector, respectively. ELISA and Western blot were used to examine the immune responses and immunoreactivity of antisera. Sperm-egg penetration assay was performed to examine the effect of anti-mCRISP1 antibodies in vitro fertilization of mouse oocytes. Fertility and mean litter size were analysed by natural mating. Histological analysis was carried out to look for potential immunopathologic effects of the antibodies. COS-7 cells transfected with pcDNA-mCRISP1 present the expression of mCRISP1. Both r-mCRISP1 and pcDNA-mCRISP1 raised an immune response against r-mCRISP1 protein and native CRISP1 in mouse sperm. The titres of anti-mCRISP1 antibodies from DNA immunized mice were significantly lower than that of r-mCRISP1 immunized mice, but it lasted relatively longer. Male and female pcDNA-mCRISP1 injected animals presented a statistically significant reduction in their fertility with no signs of immunopathologic effects. These studies demonstrated the feasibility of generating an immune response to mCRISP1 protein by DNA vaccine and pcDNA-mCRISP1 plasmid causing significant anti-fertility potential.

Comparison ofMycoplasma synoviaeisolates by immunoblotting

SDS-polyacrylamide gel electrophoresis (PAGE) and immunoblot profiles using antisera from M. synoviae F10-2AS-inoculated chickens detected heterogeneity among 10 M. synoviae isolates. This variation was primarily seen in immunoblots as a difference in the molecular size of immunoreactive proteins below 50 kDa and corresponded to an area of variability seen in SDS-PAGE. No two isolates had the same immunoblot profile below 50 kDa. Hyperimmune antiserum to a M. synoviae F10-2AS protein of 41 kDa (p41) reacted with protein(s) ranging from 41 to 48 kDa in nine of 10 isolates and strongly with one of 87 kDa in one isolate. The immunoreactivity of antiserum from inoculated chickens and the hyperimmune antiserum to p41 varied from intense to weak with the 10 M. synoviae isolates. All of the highly immunogenic proteins of M. synoviae F10-2AS, including p41, partitioned into the Triton X-114 detergent phase, indicating that they are amphiphilic in nature and integral membrane proteins. The data suggest that M. synoviae is capable of varying the molecular size of a highly immunogenic integral membrane protein.

Serological analysis of 17 baculoviruses from subgroups A and B using protein blot immunoassay

We used the protein blot radioimmunoassay technique (Smith and Summers, 1981) to evaluate the immunoreactivity of antisera made against SDS-disrupted baculovirus structural proteins. Eight antisera (six for NPVs and two for GVs) were tested against eleven nuclear polyhedrosis (NPV) and six granulosis viruses (GV). Homologous reactions revealed that SDS-disrupted baculovirus antigens elicited antibodies which reacted with several of the virus proteins. Heterologous reactions of each viral antiserum were determined with a total of 17 baculoviruses. Using these data and that of Smith and Summers (1981), we were able to detect a number of shared antigenic determinants which revealed some specific serological relationships among some of the 17 viruses tested. The MNPVs showing distinct serological relatedness included those of Autographa californica, Rachiplusia ou, Anticarsia gemmatalis, Choristoneura fumiferana, and Orgyia pseudotsugata. Heliothis armiger MNPV and Heliothis zea SNPV shared cross-reacting antigenic determinants as did Trichoplusia ni SNPV, Pseudoplusia includens SNPV, and Heliothis zea SNPV. The GVs. of T. ni, H. armiger, and Spodoptera frugiperda were also serologically related. Also reactions were detected which indicated the presence of common antigenic determinants among different baculovirus subgroups in addition to those originally detected in our earlier study (Smith and Summers, 1981). Because of the large number of structural proteins in baculoviruses and the lack of information concerning their structural or functional roles in the virus group specific antigenic determinants could not be identified. However, protein blot RIA allows specific identification of baculoviruses and the number and molecular weight of homologous and heterologous proteins sharing antigenic determinants.