Role of HLA antigens in Rh (D) alloimmunized pregnant women from Mumbai, Maharashtra, India

Immunogenetic studies in various diseases provide potential genetic markers. We have studied the incidence of HLA A, B, C, DR and DQ loci antigen in Rh (D) antigen isoimmunized mothers compared to those nonimmunized isoimmunized Rh negative mothers. Seventy six mothers who were immunized to Rh (D) antigen due to pregnancy (responders) and fifty four mothers who did not develop Rh (D) isoimmunization despite positive pregnancies (nonresponders) were selected for the study. Standard methods of serological HLA typing, ABO and Rh (D) groups, and screening for Rh D antibodies were used. 392 unrelated individuals from the population were compared as controls. In addition 45 unrelated individuals from the same population were typed for HLA DRB and DQB gene using PCR-SSP kits. The genotype frequencies of HLA A2, A3, A28, B13, B17, B35, B52, B60, Cw2, Cw6, DR4, and DQ3 were significantly increased, while the frequencies of the HLA A11, A29, A31, B7, B37, B51, Cw1 and DR9 were decreased in the responder women when compared to the non-responder women. HLA A30 (19) split antigen was not identified in immunized women while HLA A23 (9) split antigen was not identified in non immunized women. HLA A3, B17, Cw2 and DR4 showed a significant relative risk among the immunized responder women. When compared with Rh immunized women (responders) reported from USA, England and Hungary the phenotype frequencies of HLA A11, A24, A28, B5, B17, B40, DR2 and DR5 were increased while HLA A23, B8, B18, and DR6 were decreased in the Indian Rh immunized women. Two locus haplotype frequency analysis observed among the responders women revealed that among the significant haplotypes expressed A2–B5, B7–Cw1, DR2–DQ1 were highly significant haplotypes in positive linkage, while A1–B5, and A1–B7 were in significant negative linkage disequilibrium. The haplotype frequencies were ≤one when these common hapoltypes were compared with control population. Thus in the present study it is evident that the inheritance of HLA A3, B17, Cw2 and DR4 increases the relative risk factor by 2.6 times among Indian Rh isoimmunized women. Further, it is evident that there are significant differences in the observed HLA antigen frequencies and two locus haplotypes in Rh isoimmunized women when compared to women from USA, UK and Hungary due to extreme HLA polymorphism in different populations of the world     

Natural antibodies and immune antibodies of human ABO blood group system

Sera of normal and isoimmunized persons genotype OO and A₁O were fractionated to separate the three main immunoglobulin components IgG, IgM and IgA from each other. The anti-B activity was measured in every fraction and individual serum. The results indicate that the natural anti-B antibodies are of IgM molecular type, contrasting with the immune anti-B antibodies which are IgM, IgG and eventually IgA. The restriction to the IgM class and other data previously reported are discussed in relation to the origin of the natural antibodies of the human ABO blood group system.     

Serological and molecular analysis of ABO and Rh blood group chimeras

The serological examination, blood transfusion strategies and the molecular analysis to blood group chimera were conducted to demonstrate existent of chimera in blood group. The blood grouping of ABO or/and RhD, newborn red blood cells separated by capillary centrifugation. Aabsorption tests and DTT treated agglutination erythrocyte tests were implemented in four patients. Further molecular biological research was conducted on one patient''s sample. The results showed that for patient 1: ABO blood group was AB/B chimera, Rh blood cells contained the RhCE chimera gene; Patient 2: Rh blood cells contained the RhD chimera gene; Patient 3: ABO blood group was AB/B chimera, Rh blood cells contained the RhD chimera gene; Patient 4: ABO blood group was O/B chimera, Rh blood cells contained the RhCE chimera gene. The study suggests that the individuals categorized as chimeras are likely to be more common than existing literature reports. According to the serological tests, in the absence of a history of recent blood transfusion or disease to cause reduced antigen, the phenomena of hybrid aggregation of the ABO and Rh blood system were the main feature. In terms of transfusion strategy, the selection of ABO and Rh blood groups should be depended on the group of cells with more antigens.     

Incidence of Maternal Rh Immunization by ABO Compatible and Incompatible Pregnancies

The incidence of maternal Rh immunization in Rh-negative women following a single ABO compatible Rh-positive pregnancy is about 17%. This incidence was determined by following Rh-negative women through two Rh-incompatible pregnancies and analysing their sera for anti-Rh at the time of delivery of their second observed pregnancy. Maternal Rh immunization occurs almost exclusively after delivery; however, antibodies may not be detectable in the absence of further antigenic stimulation.The incidence of maternal Rh immunization when maternal-foetal ABO incompatibility is also present is 9–13% and 17% for group O and non-group O women respectively. This study emphasizes the need to offer Rh-immune prophylaxis to Rh-negative women having Rh-positive infants whether or not ABO incompatibility exists between the mother and infant.     

Genetic polymorphisms in juvenile-onset diabetes

Nine genetic polymorphic systems (ACP1, PGM1, ADA, AK, G-6-PD, Hp, ABO, Rh, MN), were studied in a series of 138 subjects affected by JOD. Differences between diabetic patients and controls were observed in the distribution of phenotypes of the red cell acid phosphatase (ACP1), and the ABO and MN blood groups.     

Over-expression and secretion of angiogenin in intrauterine growth retardation placenta

Human angiogenin is a potent inducer of neovascularization. There is a strong evidence to suggest that it might be involved in morphological and angiogenic changes in the placenta, that are necessary for a successful fetal outcome during pregnancy. However, its precise role in the pathogenesis of abnormal pregnancies is yet unknown. Intrauterine growth retardation (IUGR), an abnormal pregnancy is not a specific disease entity per se, but rather a manifestation of many possible fetal and maternal disorders. In this study, we demonstrated, for the first time, that placental explants in vitro secrete significantly elevated levels of angiogenin in placental tissues from patients with IUGR. We also observed enhanced mRNA expression in placenta from these patients. In addition, using the immunohistochemical methods, we observed identical staining of angiogenin to villous syncytiotrophobalst and fetal endothelial cells in both IUGR and normal placenta. Functionally active placental explants were used to detect immunoreactive angiogenin in conditioned media of all the samples from IUGR placenta and normal term group. The mean levels of angiogenin secreted by IUGR placenta were 1.4-, 1.6-, and 1.3-fold higher (P < 0.01) than normal term samples at 24, 48, and 72 hr of culture, respectively. Expression profiles of angiogenin from term and IUGR cases are in agreement with its mRNA levels and immunoblot analysis. In conclusion, the significant elevated levels of angiogenin in IUGR placenta may provide a molecular mechanism for the abnormal placental development.     

Segregation distortions of the ABO and Rh systems in malformed newborns

This study compares the segregation of the ABO and Rh systems between malformed newborns and a control group with two purposes: (1) to evaluate the participation of genetic factors associated with these blood groups in the production of congenital malformations, and (2) to prove, indirectly, the existence of reproductive losses associated with congenital malformations. The newborns and their mothers were typed for ABO and Rh groups. Gene frequencies were similar in malformed and control newborns. In the female malformed newborns, an excess of O-B pairs and a deficit of B-B pairs in the ABO system, and an excess of Rh(-)-Rh- pairs in the Rh system were found.     

ABO and Rh blood groups in diabetes mellitus

The present investigation was conducted with a view to testing the hypothesis that there is some association between blood groups (ABO and Rh) and diabetes mellitus. 520 proven cases of adult diabetes mellitus from the Diabetic Clinic of Rajendra Hospital, Patiala, were studied in 1979-1980. A large sample of 6204 normal individuals studied by Jolly et al. (1969) for ABO and Rh blood groups was taken as control for comparison with the patients. There is a strong indication of an association of diabetes mellitus with blood groups, especially with A, AB and Rh-positive blood groups. The maximum differences are in the AB groups in the two series and minimum in the A group. Individuals with gene p seem to be more susceptible to this disease. Thus the association between blood groups and diabetes mellitus is not a chance finding, but implies an aetiological relationship.     

Frequencies of ABO, Rh and Kell phenotypes in couples from Han, Kazak, Uyghur and Hui in Xinjiang: an inheritance simulation model for blood group incompatibility in new-born

The Xinjiang region with residents from more than 13 minorities represents an area of many diverse ethnicities. This ethnic diversity in relation to their blood groups and immune status may have a consequential impact on the clinical status of married couples. To evaluate the risks of haemolytic disease in new-born infants, we investigated the rate of blood-group incompatibility among 487 married couples from four ethnic minorities, namely the Han, Hui, Uyghur and Kazak populations. Han minority married couples showed significantly different ABO, Rh and K phenotype frequencies between marrial relationship, whereas there was no significant difference in ABO, Rh and K phenotypes between the Uyghur, Hui and Kazak .There was a significant difference between ABO blood types in Han married couples, in the Kazak Rh-C phenotype and in the Uyghur Rh-D phenotype. The Hui married couples only demonstrated ABO, Rh and K phenotypes. The Hui minority showed the highest incompatibility rate for Rh-C and Rh-E phenotypes between mothers and their new-born infants. The highest incompatibility rate for the ABO phenotype occurred in the Kazak group. These results particularly demonstrate the clinical issues relating to ABO and Rh incompatibility, in the Kazak and Hui minorities, respectively.     

全自动血型仪的应用评价

目的了解全自动血型鉴定系统用于检测ABO血型和Rh(D)血型的效果。方法采用深圳爱康AK03A型数字血型仪对北海市中心血站2010年5月—2011年12月无偿献血者20702份样本进行ABO血型和Rh(D)血型鉴定,并与手工试管法进行对比验证。结果发现正反定型不一致的有75例,ABO初筛血型错误的36例,Rh(D)阴性的53例。结论全自动血型鉴定系统检测血型快捷、方便、高效,能降低检测人员的工作强度,但判读结果时需要与手工试管法结合进行验证,才能得出正确的结果。     

Blood group typing among Brazilian brown-howlers (Alouatta fusca)

An attempt was made to investigate ABO, Rh and MN blood antigens of 108 monkeys belonging to the species Alouatta fusca (brown-howlers).Data concerning ABO antigens did not reveal any positive blood sample for anti-H and anti-A₁ sera.In relation to the Rh system, anti-D, anti-c, anti-C and anti-E sera were used and a different pattern of antigen response was apparent. Positive blood samples were detected only through an indirect Coomb's test among c. 50% of the subjects examined for anti-D serum; and 100% for anti-c serum. For anti-E serum, c. 40% and for anti-C serum c. 50% were positive.The serological response for anti-M and anti-N sera was invariably negative as it was also found out amongst monkeys in the New World.Several techniques were employed to demonstrate the similarity between A and B antigens of monkeys and human erythrocytes and the results indicate the presence of a non-specific antigen in the erythrocyte and a corresponding antibody in the serum of the monkeys.Brown-howler erythrocytes were agglutinated by human serum corresponding to all four ABO blood types. Conversely, the hour types of human erythrocytes were agglutinated by serum belonging to any type of brown-howler subject.It was not possible to investigate antigens of the saliva from monkeys and on attempt to detect these antigens in kidney extract had no success.As a general picture, it was concluded that, corresponding to human ABO blood types, two systems were apparent, one including two antigens and the other just one.     

Synthesis of human Placental Lactogen in Xenopus oocytes

Oocytes from Xenopus laevis were injected with polysomes from normal human term placenta.Synthesis of the protein hormone Human Placental Lactogen (HPL) in the oocytes was demonstrated by specific immunoprecipitation with anti-HPL serum.Analysis of the immunoprecipitates on SDS-polyacrylamide gel revealed one peak, with a migration distance corresponding exactly to that of [¹⁴C]-Radioacetylated HPL added as a marker.Two days after injection the mRNA was still able to direct the synthesis of HPL.     

Acuity of selective mechanisms operating on ABO, Rh, and MN blood groups

Selection in ABO, Rh, and MN blood groups was studied in 216 matings and their children in an endogamous population. Incompatibility status with respect to these three systems was considered simultaneously. There is no effect of incompatibility on number of pregnancies. Analysis of variance between groups confirms that prenatal loss is associated with incompatibility, and it is greater when the matings are incompatible for any two systems. There is no significant intergenerational change in ABO and Rh polymorphisms. Segregation analysis for the ABO system suggests that there is no significant difference in the proportion of A, B, and O children, based on the compatibility of the parents, while analysis for Rh-D system showed a segregation distortion which is not related to the known antigenic specificities (mother-child incompatibility).     

Transrectal combined thickness of the uterus and placenta in normal pregnant Egyptian buffalo-cows

The combined thickness of the uterus and placenta (CTUP) is one of the characteristics that can be used to assess fetal development and/or placental function in bovine. The current study was designed to establish reference values for the CTUP throughout pregnancy in normal pregnant buffalo-cows. The CTUP at the intracotyledonary space was measured monthly from the second month until full term using electronic calipers of the ultrasound machine. The CTUP increased monthly from 2.5 mm at the second month to 12 mm at the full term. During the last trimester, the monthly increase in the CTUP was higher than that recorded during the first and second trimesters. The result of the current study can be used as normal values for future studies of CTUP in pathologically pregnant buffalo-cows.     

Rh isoimmunization,Manitoba, 1963-75.

The number of Rh-isoimmunized pregnancies in Manitoba has been reduced from 223 and 228 in the years ending Oct. 31, 1963 and 1964 to 60 and 62 in the years ending Oct. 31, 1974 and 1975. The number per 1000 total births in the same years has decreased from 10.0 and 10.6 to 3.4 and 3.5 Perinatal mortality rates in those years decreased from 13.8 amd 15.7% to 0 and 2.2%, respectively. The number of perinatal deaths has been reduced from 55 in the first 2 years reported to 1 in the last 2 years. Among the 121 isoimmunized women pregnant in the 2-year period ending Oct. 31, 1975, isoimmunization was due to failure to give Rh immune globulin after delivery in 33 and failure to give it during pregnancy in 48. Of the remaining 40, 37 were immunized before Rh immune globulin became available. Complete prevention of Rh isoimmunization and therefore of all perinatal deaths from Rh erythroblastosis can only be achieved through universal Rh testing prenatally and immediately after delivery, and institution of an antenatal Rh prophylaxis program.     

Blood groups and HLA antigens in patients with abdominal aortic aneurysms

Frequencies of blood groups (ABO, Rh, MNSs, P, Kell, Lewis and Duffy) and HLA antigens were studied in a series of patients from northern Sweden with abdominal aortic aneurysms. The following significant differences from the controls were found: a decreased frequency of the Rh-negative blood group and increased frequencies of the Kell-positive and MN blood groups. Previously reported associations with the ABO and Rh systems were not confirmed.     

Cloning and characterization of an apoptosis-associated gene in the human placenta

Placenta is a transient feto-maternal association that develops during mammalian pregnancies. Human placental tissue during the first trimester of pregnancy is an actively dividing and differentiating tissue, while near term, it represents a fully differentiated unit performing many life-sustaining functions for the fetus. Previous studies have demonstrated that the percentage of placental cells that undergo apoptosis is greater at full term as compared to the first trimester of pregnancy. In this study, we undertook a study aimed at gaining an insight into the kind of genes expressed in the two developmentally distinct stages of gestation ie, the first trimester and term using Differential Display RT-PCR. Cloning and sequencing of one of the differentially expressed cDNAs from term placental tissue revealed that it is a novel gene, referred to as T-18 in the text. In this study, we also examined the regulation of this gene during apoptosis in the human placenta. A model for analysis of placental apoptosis was established by incubating placental villi in serum-free culture medium. It was observed that apoptosis occurred rapidly following incubation of placental villi without tropic support, and the proposed free-radical scavenger, superoxide dismutase (SOD) suppressed apoptosis in the placenta. Interestingly, the levels of T-18 mRNA increased significantly during spontaneous induction of apoptosis and decreased when apoptosis was blocked by SOD. These data clearly suggest that there is a strong correlation between the expression of T-18 and placental apoptosis and that T-18, may play a significant role in this process. Furthermore, the establishment of a defined in vitro explant culture model should facilitate elucidation of factors, which regulate apoptosis in human placenta.     

Antenatal prophylaxis of Rh isoimmunization: 28-weeks'-gestation service program

Two (0.18%) of 1086 Rh-negative primigravidas or multigravidas treated similarly in all previous pregnancies, who were given a single injection of Rh immune globulin (300 μg) at 28 weeks'' gestation and subsequently were delivered of Rh-positive babies, had demonstrable Rh isoimmunization at the time of that injection and must be considered “logistic” failures of antenatal prophylaxis. The remaining 1084 (who were treated again after delivery) had no evidence of Rh isoimmunization at delivery and none of the 512 screened at 6 months after delivery appeared to be immunized. If the 28th-week injection had not been protective, one would have expected 14 of the 1084 to have been demonstrably Rh isoimmunized and evidence of Rh isoimmunization to have persisted in 6 of the 512 observed 6 months after delivery.Six of 719 Rh-negative multigravidas who had not received Rh immune globulin after previous pregnancies or had been treated only after delivery showed evidence of Rh isoimmunization despite a single injection of Rh immune globulin at 28 weeks in a subsequent pregnancy. In three of the six the cause was most likely “sensibilization” due to previous exposure to Rh-positive blood or an untreated Rh-positive pregnancy. in 3 of the remaining 716 (0.42%) there may have been true failure of antenatal Rh prophylaxis administered at the 28th week. One would have expected this figure to be 12 of 716 if antenatal Rh prophylaxis at 28 weeks'' gestation were totally unsuccessful.It is concluded that a single intramuscular injection of Rh immune globulin, 300 μg, is 88% effective in preventing Rh isoimmunization during pregnancy in Rh-negative primigravidas and in multigravidas treated antenatally in all previous pregnancies, and is 75% effective in preventing Rh isoimmunization in Rh-negative multigravidas untreated during previous pregnancies. The majority of failures are due to Rh isoimmunization during pregnancy prior to antenatal prophylaxis at 28 weeks.     

Demonstration of the activity of a rhesus-positive antigen in the erythrocytes of rhesus-negative donors and an increase in the expression of ABO system antigens after ultraviolet irradiation of the blood

UV irradiation (254 nm) in doses increasing erythrocyte (Er) hemolysis by 5 to 32% was found to stimulate the agglutination activity of ABO and Rhesus (Rh) system antigens. The stimulation effect was the higher the lower the antigen activity before irradiation. In the Rh-negative (Rh-)-Er, irradiation induced the Rh0(D)-like antigen specific activity suggesting that this antigen may be present in the Rh--Er membrane. Expression of Rh0(D)-antigen in Rh--Er, stimulation of its activity in Rh-positive cells, and activation of ABO system antigens may result from photo-chemical destruction of the outer membranous layer of the ER.