A direct blood polymerase chain reaction approach for the determination of GP.Mur (Mi.III) and other Hil+ Miltenberger glycophorin variants.

BACKGROUND: GP.Mur (Mi.III) is a glycophorin B-A-B hybrid sialoglycoprotein expressing several potent immunogens, including Mi(a), Mur, and Hil. GP.Mur is considered one of the most important red blood cell (RBC) phenotypes in blood banking in Southeast Asia. However, there are no antibodies commercially available for the screening of GP.Mur RBCs. STUDY DESIGN AND METHODS: To develop a direct blood polymerase chain reaction (PCR) approach for the screening of GP.Mur cells, we first confirmed the genomic sequence differences among four GP.Mur and three Mi(a-) samples by sequencing their GYP.Mur and GYPB genes. With these data, we designed PCR primers that best discriminate GYPB and GYP.Mur. Our primer design also allows the detection of other Hil+ glycophorin variants. We also constructed two plasmids--pGBi2i3 and pMiIIIi2i3--which serve as the negative and positive control DNA, respectively, for the PCR procedure. Additionally, we designed a control PCR to be run side by side with the typing PCR. RESULTS: Because of the high specificity of our primers, we found it unnecessary to extract DNA from blood samples for PCR. We have tested this PCR method on 379 fresh and frozen blood samples. The results were further validated by serology and DNA sequencing and were shown to be completely accurate in our hand. We also found that the rapid genotyping method--high-resolution melting--can be a timesaving alternative for DNA sequencing. CONCLUSION: This direct blood PCR approach for determination of GP.Mur and related Hil+ phenotypes is reliable and economical and is expected to be useful for blood banking in Southeast Asia.

Composition of erythrocytic (ABO, Rh-Hr, Kell, MN) group antigens characteristic of the Ozurgeti district's population.

Erythrocytic group antigens represent a genetically stably determined trait. Investigation of antigens of the said system in different regions is of the greatest importance in terms of both the creation of demographic data of the region as well as practical medicine, especially for transplantology and transfusiology. The peripheral or venous blood of 232 local natives (healthy donors) of Ozurgeti district of Guria region has been taken as the test subject. The test subject was taken by random methods in different vilifies (Bakhvi, Mshvidobauri, Ozurgeti, Likhauri, Gurianta, Bokhvauri, Dvadzu, Pampaleti) To identify the ABO, Rh-Hr, Kell, MN system antigens, an express-method using monoclonal antibodies has been applied. In studying the ABO system, it was fixed that the highest distribution frequency was characteristic of the 0(I) group (52.3±3.2%), then follows the group A(II) (38.5±3.2%). The distribution frequency of the B(III) group is (8.2±1.8%) and that of AB(IV)--(0.8±0.5). The population's 85.2±2.32% is the carrier of the Rh+ phenotypic group, while 14.7±2.3% belongs to the Rh-phenotypic group. In studying the concentration of alleles, the low concentration of p(K) allele was detected that equaled 0.2; the concentration of q(K) allele made 0.8, that of p(M)--0.65, and that of q (N) - 035.

Grupos sanguíneos y enfermedad / Blood groups and disease

La membrana eritrocitaria sirvió como modelo general para el conocimiento de la membrana plasmática. Algunas de sus estructuras son antígenos pertenecientes a los sistemas de grupos sanguíneos y están siendo caracterizadas molecular y funcionalmente como receptores, transportadores o enzimas, incluso como puertas de entrada para patógenos. Así, el Plasmodium vivax (causante de la malaria) requiere la glucoproteína Duffy para penetrar en el interior de los hematíes humanos, y el antígeno principal del sistema P (P1) es también el receptor para el acceso del parvovirus B19. Estos antígenos no siempre se limitan a los glóbulos rojos, sino que pueden influir en diversos tejidos, el plasma o las secreciones con importantes relaciones patogénicas. Ciertas cepas agresivas de Eschirichia coli precisan antígeno P1 para anclarse al epitelio urinario, el antígeno Lewis(b) es el receptor de Helicobacter pylori en la mucosa gástrica, el anti-B de los sujetos con los grupos sanguíneos O y A podría ayudarles a combatir las bacteriemias por E. coli, el grupo Lewis condiciona las concentraciones séricas de CA-19.9 y el efecto protector de la leche materna. Sin embargo, la principal influencia sería la hipocoagulabilidad observada en la población de grupo O (valores inferiores de factor VIII) asociada con una prevalencia menor de enfermedades tromboembólicas

The erythrocyte membrane was used as general model for the plasma membrane knowledge. Some of their structures are antigens from blood group systems being characterized at molecular and functional level as specific receptors, transporters or enzymes, even receptors for infectious agents. Plasmodium vivax malarial parasites require the Duffy blood group glycoprotein to penetrate into human red blood cells and the main antigen of P system (P1) is also the Parvovirus B19 receptor. Furthermore, these substances have an effect on several tissues, plasma and secretions involving pathogenic relationships. Certain aggressive Escherichia coli strains require the P1 antigen to attach to the urothelial cells, the Lewis(b) antigen is the gastric receptor for H. pylori, the anti-B from O or A individuals might protect them against the sepsis produced by E. coli, the Lewis group determines the CA-19.9 serum levels or the protective effect of breast milk. However, the most important effect could be the plasma hypocoagulability observed among the O blood group population (with lower factor VIII levels) in association with a reduced prevalence of thromboembolic diseases

[The identification of antigens of systems AB0, GM, and haptoglobin (HP) phenotypes in mixed bloodstains of human and domestic goose]. / Identifikatsiia antigenov sistem AB0, GM and fenotipov gaptoglobina (HP) v smeshannykh piatnakh krovi cheloveka i domashnikh gusei

The traditional methods of investigations according to systems AB0, Gm and Hp were used to define the serological specificity of home gooses' blood. The experimental examinations' results related with mixed bloodstains of man and home gooses are described. A possibility is demonstrated to identify the blood group factors of man in bloodstains with admixture of home-goose blood.

Expression of the MN antigen in cervical papanicolaou smears is an early diagnostic biomarker of cervical dysplasia.

A new tumor-associated antigen, MN, has been shown to be expressed in virtually all cervical carcinomas and the majority of cervical intraepithelial neoplasia, but not in normal cervices (S. Y. Liao et al., Am. J. Pathol., 145: 598-609, 1994). Therefore, we postulated that the exfoliative cells in cervical Papanicolaou (Pap) smears would reflect the MN immunoreactivity seen in the tissue sections, and high levels of MN expression in the exfoliative cells would indicate the presence of dysplasia in the cervix. A total of 305 cervical Pap smears, with histological confirmation, representing all categories of the Bethesda System, were immunohistologically examined. We found that high levels of MN expression in exfoliative cells were not restricted to the dysplastic cells but were observed also in the normal endocervical cells (NECs) when dysplasia was present in the tissue biopsies. Overall, the rates of positive MN immunostaining of the dysplastic cells in low- and high-grade squamous intraepithelial lesions and invasive carcinoma were 35 (65%) of 54, 44 (77%) of 57, and 12 (92%) of 13, respectively. However, diffuse MN immunoreactivity of the atypical and/or dysplastic endocervical columnar cells was seen in all cases (100%) of adenocarcinoma in situ (AIS; n = 23) and adenocarcinomas (n = 8). In the groups with cytological diagnoses of atypical squamous cells or atypical glandular cells of undetermined significance (ASCUS and AGUS, respectively), MN positivity was seen in 47% of ASCUS (22/47) and 55% of AGUS (12/22). Dysplastic tissues were identified in all MN-positive cases. In contrast, all MN-negative atypical Pap smears were confirmed histologically to be benign cervix with one exception, in which the cytological diagnosis was ASCUS and focal low-grade squamous intraepithelial lesions were found in the cervix. The study also included 89 cases with cytological diagnoses of within normal limits/benign cellular changes. Among these, 10 Pap smears expressed diffuse MN antigen in the NEC, and dysplasia (8 cases of low-grade squamous intraepithelial lesions, 2 AIS) was found in the cervices. None of MN-negative cases with "within normal limits" cytology contained dysplastic cervices. Therefore, it would seem that diffuse MN antigen expression in the NEC may be an indicator of cervical dysplasia. Thus, MN antigen might serve as an early biomarker of cervical neoplasia. The combination of detection via cytology and MN immunostaining resulted in no false negatives and also discriminated between cellular atypia due to benign reactive changes versus cellular atypia due to dysplasia in the category of ASCUS and AGUS. In particular, it was found in the AGUS group that diffuse MN immunostaining restricted to atypical columnar cells was diagnostic for AIS. These findings indicate that MN antigen expression is an important diagnostic biomarker of glandular neoplasia and a valuable adjunct to cytological diagnosis of ASCUS and AGUS.

Asociación de marcadores eritrocitarios (ABO, MNSs, Rh, Lewis, P1) con recurrencias y anomalías anatómicas y funcionales en niños con infección del tracto urinario / Association of erythrocyt markers (ABO, MNSs, Rh, Lewis, P1) with recurrences and anatomical-functional abnormalities in children with urinary tract infection

Para contribuir a la identificación de niños con infección del tracto urinario con mayor riesgo de ubicación alta o baja, recurrencias y alteraciones radiológicas o ultrasonográficas (complicaciones) se estudiaron las asociaciones entre éstas y la distribución de marcadores eritrocitarios (ABO, MNSs, Rh, Lewis, P1) en 309 casos de infección urinaria. No se encontró asociación entre algún polimorfismo eritrocitario en particular con las mencionadas categorías, pero si entre el fenotipo P1 y la etiología Escherichia coli (OR=3,07; IC 95 porciento=1,13 a 8,6; p<0,02) y la ausencia de etiología no E. coli con el fenotipo B+(0/26) sin llegar a niveles de significación. Estos hallazgos sugieren que en niños con infección urinaria, estos fenotipos, por separado, probablemente tienen acciones independientes y aditivas

The differences in significance of alpha 2,3Gal-linked and alpha 2,6GalNAc-linked sialic acid residues in blood group M- and N-related epitopes recognized by various monoclonal antibodies.

The blood group M and N determinants of glycophorin A (GPA) contain O-linked oligosaccharide chains with alpha 2,3Gal-linked and alpha 2,6GalNAc-linked sialic acid residues which are required for the activity of most epitopes recognized by various anti-M and anti-N antibodies. In order to check whether these two types of sialic acid residues differ in their contribution to antigenic properties, the GPA-M and GPA-N preparations with monosialylated oligosaccharide chains were obtained and tested for binding of anti-M and anti-N monoclonal antibodies (MAbs). The GPAs with sialic acid residues linked to Gal (GPA2,3) were obtained by selective resialylation of asialoGPAs with alpha 2,3-sialyl-transferase. These preparations were tested by inhibition of binding of MAbs to enzyme-linked immunosorbent assay (ELISA) plates coated with the respective untreated target antigens. The GPAs with sialic acid residues linked to GalNAc (GPA2,6) were generated by treating GPAs adsorbed on ELISA plates with Newcastle disease virus (NDV) isolate (expressing sialidase specific for alpha 2,3Gal linkage), which was followed by testing the binding of MAbs to NDV-treated antigens. Different patterns of activity were obtained among 14 MAbs specific for sialic acid-dependent epitopes (eight anti-M and six anti-N). The results indicated that at least half of the MAbs showed distinct requirements for the presence of only one of two kinds of sialic acid residues (Gal or GalNAc linked) in the epitope. Only four MAbs (two anti-M and two anti-N) did not react with any of the 'monosialylated' forms of GPA.(ABSTRACT TRUNCATED AT 250 WORDS)