Histo-blood group antigens and rotavirus vaccine shedding in Nicaraguan infants.

ABO, Lewis and secretor histo-blood group antigens (HBGA) are susceptibility factors for rotavirus in a P-genotype dependent manner and can influence IgA seroconversion rates following rotavirus vaccination. To investigate the association between HBGA phenotypes and rotavirus vaccine shedding fecal samples (n = 304) from a total of 141 infants vaccinated with Rotarix (n = 71) and RotaTeq (n = 70) were prospectively sampled in three time frames (≤3, 4-7 and ≥8 days) after first vaccination dose. Rotavirus was detected with qPCR and genotypes determined by G/P multiplex PCR and/or sequencing. HBGAs were determined by hemagglutination and saliva based ELISA. Low shedding rates were observed, with slightly more children vaccinated with RotaTeq (19%) than Rotarix (11%) shedding rotavirus at ≥4 days post vaccination (DPV). At ≥4 DPV no infant of Lewis A (n = 6) or nonsecretor (n = 9) phenotype in the Rotarix cohort shed rotavirus; the same observation was made for Lewis A infants (n = 7) in the RotaTeq cohort. Putative in-vivo gene reassortment among RotaTeq strains occurred, yielding mainly G1P[8] strains. The bovine derived P[5] genotype included in RotaTeq was able to replicate and be shed at long time frames (>13 DPV). The results of this study are consistent with that HBGA phenotype influences vaccine strain shedding as similarly observed for natural infections. Due to the low overall shedding rates observed, additional studies are however warranted.

Breast cancer humoral immune response: involvement of Lewis y through the detection of circulating immune complexes and association with Mucin 1 (MUC1).

BACKGROUND: In cancer patients, MUC1 glycoprotein may carry Lewis y which could be involved in immune response. PURPOSES: 1- to evaluate the presence of Lewis y and MUC1 in circulating immune complexes (Lewis y/CIC and MUC1/CIC, respectively) and their correlation; 2- to analyze the possible presence of Lewis y in carbohydrate chains of tumoral MUC1 glycoprotein and 3- to correlate serum and tissue parameters considered. METHODS: Pretreatment serum and tissue breast samples from 76 adenocarcinoma, 34 benign and 36 normal specimens were analyzed. Anti-MUC1 and anti-Lewis y MAbs were employed. To detect Lewis y/CIC and MUC1/CIC, ELISA tests were developed; serum samples containing MUC1 were previously selected by Cancer Associated Serum Antigen (CASA). Immunoprecipitation (IP) was performed in 9 malignant, benign and normal samples and analyzed by SDS-PAGE and Western blot. Lewis y and MUC1 expression was studied by immunohistochemistry (IHC). Statistical analysis was performed employing principal component analysis (PCA), ANOVA, Tukey HSD, Chi square test and classical correlation (p < 0.05). RESULTS: By ELISA, Lewis y/IgM/CIC levels showed statistically significant differences between breast cancer versus benign and normal samples; mean +/- SD values expressed in OD units were: 0.525 +/- 0.304; 0.968 +/- 0.482 and 0.928 +/- 0.447, for breast cancer, benign disease and normal samples, respectively, p < 0.05. Lewis y/IgG/CIC did not show any statistically significant difference. MUC1/IgM/CIC correlated with Lewis y/IgM/CIC. By CASA, 9 samples with MUC1 values above the cut off were selected and IP was performed, followed by SDS-PAGE and Western blot; bands at 200 kDa were obtained with each MAb in all the samples. By IHC, with C14 MAb, 47.5%, 31% and 35% of malignant, benign and normal samples, respectively, showed positive reaction while all the samples were positive with anti-MUC1 MAb; in both cases, with a different pattern of expression between malignant and non malignant samples. CONCLUSION: Our findings support that in breast cancer there was a limited humoral immune response through Lewis y/IgM/CIC levels detection which correlated with MUC1/IgM/CIC. We also found that Lewis y might be part of circulating MUC1 glycoform structure and also that Lewis y/CIC did not correlate with Lewis y expression.

Fetal hydrops and other variables associated with the fetal hematocrit decrease after the first intrauterine transfusion for red cell alloimmunization.

OBJECTIVE: To evaluate the influence of fetal hydrops and other variables on fetal hematocrit (Hct) decrease after the first intrauterine transfusion (IUT) in alloimmunized pregnancies. METHODS: From 1996 to 2006, the data of all alloimmunized pregnancies submitted to IUT were assessed. Exclusion criteria included: fetuses submitted to intraperitoneal transfusion; pregnancies complicated by other fetal abnormalities; pregnancies submitted to only one IUT, and cases in which posttransfusion or pretransfusion blood samples were not obtained. Linear regression models were implemented to assess the relationship between the rate of Hct fall after the first IUT and the following variables: fetal hydrops; antibody titer; gestational age at the first IUT; number of days between the first and second IUT; pretransfusion and posttransfusion fetal Hct values. RESULTS: Fifty fetuses fulfilled the study criteria. The fetal Hct decrease after the first IUT was 1.21 (range 0.18-2.3) %/day. The variables independently associated with the fetal Hct drop after the first IUT were the fetal hydrops (p = 0.000), the pretransfusion fetal Hct (p = 0.001) and the posttransfusion fetal Hct (p = 0.016). CONCLUSION: Fetal hydrops, pretransfusion fetal Hct and posttransfusion fetal Hct seem to influence the fetal Hct decrease between the first and second IUT. These findings may be helpful for estimating the rate of fetal Hct drop and programming the following IUT.

Anti-Lewis alloimmunization: report of seven cases.

OBJECTIVE: The purpose of this study was to investigate the perinatal results of seven pregnant women with anti-Lewis antibodies and evaluate the need to screen for these antigens during routine prenatal care. SETTING: São Paulo Universtity Hospital, São Paulo, Brazil. POPULATION: 200 Rh-negative pregnant women with a positive indirect Coombs test, managed during a 6-year period. METHODS: The charts of all patients were reviewed to collect pertinent data and the variables were analyzed. MAIN OUTCOME MEASURES: Indirect Coombs test titer, intrauterine transfusion, mode of delivery, gestational age at birth, birthweight, neonatal transfusion, duration of neonatal hospitalization and perinatal mortality. RESULTS: All newborn infants were classified as adequate for gestational age at birth and none needed intrauterine or neonatal transfusions. All infants, except one, were discharged in good health on the third day after birth. CONCLUSIONS: Alloimmunized pregnancies (Levis antigens) have good perinatal results.

Colorectal carcinoma: a pathologic and immunopathologic study.

Lewis-related blood group antigens are carbohydrate determinants carried on surface glycoproteins and glycolipids of erythrocytes and some epithelial cells. Their expressions in colorectal mucosa has been considered either as a marker of differentiation antigens or of tumor associated antigens. We prospectively studied Lewis-related blood group antigens in patients undergoing colectomy for colonic adenocarcinoma. Immunohematological studies were performed on preoperative blood samples and ABO and Lewis blood groups were determined. Tissue samples were obtained from the adenocarcinoma and from the adjacent non-neoplastic mucosa. The degree of tumor differentiation (grade of the tumor) was analyzed on hematoxylin and eosin stained slides. Immunoperoxidase staining was performed using antibodies against A, B, Le(a), Le(b), Le(x), and Le(y) antigens. The staining results of the colonic carcinoma were compared with those of the non-neoplastic mucosa and with the patient's red blood cell Lewis phenotype. A total of 22 patients were studied, 12 males and 10 females, with an average age of 64 +/- 12 years. There were 12 modified Dukes B and 10 modified Dukes C carcinomas. Le(a), a marker of differentiation antigens, was expressed in the non-neoplastic mucosa of 82% of the specimens. Le(y), a marker of tumor associated antigens, was detected in 77% of the carcinomas while expressed only in 18% of the adjacent non-neoplastic mucosa. Le(x), supposedly a marker for tumor aggressiveness, was found in 55% of the carcinomas. However, it was virtually absent in the five carcinomas which did not invade the pericolic tissues.

Colorectal carcinoma: a pathologic and immunopathologic study

Lewis-related blood group antigens are carbohydrate determinants carried on surface glycoproteins and glycolipids of erythrocytes and some epithelial cells. Their expressions in colorectal mucosa has been considered either as a marker of differentiation antigens or of tumor associated antigens. We prospectively studied Lewis-related blood group antigens in patients undergoing colectomy for colonic adenocarcinoma. Immunohematological studies were performed on preoperative blood samples and ABO and Lewis blood groups were determined. Tissue samples were obtained from the adenocarcinoma and from the adjacent non-neoplastic mucosa. The degree of tumor differentiation (grade of the tumor) was analyzed on hematoxylin and eosin stained slides. Immunoperoxidase staining was performed using antibodies against A, B, Le(a), Le(b), Le(x), and Le(y) antigens. The staining results of the colonic carcinoma were compared with those of the non-neoplastic mucosa and with the patient's red blood cell Lewis phenotype. A total of 22 patients were studied, 12 males and 10 females, with an average age of 64 +/- 12 years. There were 12 modified Dukes B and 10 modified Dukes C carcinomas. Le(a), a marker of differentiation antigens, was expressed in the non-neoplastic mucosa of 82 per cent of the specimens. Le(y), a marker of tumor associated antigens, was detected in 77 per cent of the carcinomas while expressed only in 18 per cent of the adjacent non-neoplastic mucosa. Le(x), supposedly a marker for tumor aggressiveness, was found in 55 per cent of the carcinomas. However, it was virtually absent in the five carcinomas which did not invade the pericolic tissues