Alloimmune neonatal neutropenia in Croatia during the 1998-2008 period.

PROBLEM: The aim of this study was to estimate the incidence of the disease and to analyze laboratory data of 23 newborns undergoing serologic testing for alloimmune neonatal neutropenia (ANN) during the 1998-2008 period in Croatia. METHOD OF STUDY: Laboratory data on 23 newborns undergoing serologic testing for ANN during the 1998-2008 period and epidemiologic data on the number of live births in Croatia were analyzed. Laboratory testing for ANN included serologic screening of maternal and neonatal sera and granulocytes (neutrophils) by immunofluorescence (IF) method. The monoclonal antibody immobilization of neutrophil antigens (MAINA) was employed to determine anti-HNA antibody specificity. RESULTS: Anti-HNA antibodies were detected in seven (54%) of 13 cases of serologically positive ANN. Only anti-HLA class I antibodies were demonstrated in four (31%) of 13 cases In the 2007-2008 period of prospective data collection, the number of serologically verified ANN cases was one case per 17,323 live births. Results of the prospective study conducted at Maternity Ward, Department of Gynecology and Obstetrics, Sestre milosrdnice University Hospital Center yielded the ANN incidence of one case per 2843 live births. CONCLUSION: Monitoring of neutrophil count in neonatal blood and serologic testing for ANN in case of isolated neutropenia in the newborn contributed considerably to timely detection of ANN. DESCRIPTORS: Neonatal alloimmune neutropenia-incidence, serologic diagnosis, antineutrophil antibodies, anti-HNA, anti-HLA class I, Croatia.

The frequencies of human neutrophil alloantigens among the Japanese population.

Human neutrophil antigens (HNAs) play an important role in a variety of clinical conditions including immune-mediated neutropenia, non-hemolytic transfusion reactions, and transfusion-related acute lung injury. The aim of this study was to investigate the frequency distribution of HNAs-1 to -5 among the Japanese population. We analyzed samples from 570 healthy Japanese by molecular and serologic techniques to estimate the gene frequencies of HNAs-1 to -5. DNA samples were obtained and typed for the HNA-1 (n = 523), -3 (n = 570), -4 (n = 570), and -5 (n = 508), by molecular techniques. The HNA-1 genotype was determined by using a commercial polymerase chain reaction-reverse sequence-specific oligonucleotide probes (PCR-rSSOP) kit. The HNA-3 to -5 genotypes were determined by the PCR-sequence specific primer (PCR-SSP), previously described, with a small modification. The HNA-2a phenotype was determined in 301 donors by granulocyte immunofluorescence test. In Japanese, the gene frequencies of HNA-1a, -1b, and -1c were 0.623, 0.377, and 0.000, respectively. The frequency of HNA-2a phenotype was 0.987, and the gene frequencies of HNA-3a and -3b were 0.654 and 0.346, respectively. HNA-4a and -4b were found at 1.000 and 0.000, respectively, and HNA-5a and -5b at 0.840 and 0.160, respectively. We describe, for the first time, the frequencies of all HNAs (HNA-1 to -5) among the Japanese population. This study will be helpful for the prediction of the risk of alloimmunization to HNA, especially to determine the risk of HNA alloantibody production by transfusion of HNA incompatible blood and feto-maternal incompatibility.

Granulocyte serology: current concepts and clinical signifcance.

Applying serologic procedures to the detection of RBC and lymphocyte antigens has facilitated the identification of granulocyte antigens with established clinical significance, which are now classified in the human neutrophil antigen system. Granulocyte alloantibodies and autoantibodies have been implicated in a variety of clinical conditions including alloimmune neutropenia, autoimmune neutropenia, febrile and severe pulmonary transfusion reactions, drug-induced neutropenia, refractoriness to granulocyte transfusions, and immune neutropenia after hematopoietic stem cell transplantation. Although the intrinsically fragile nature of granulocytes contributes to the inherent challenges of granulocyte serology, several advances in laboratory procedures have improved detection of granulocyte antibodies. This review will provide a current perspective about the importance and use of granulocyte serology for detection of granulocyte antibodies that have significant medical effects.

Molecular nature of antigens implicated in immune neutropenias.

Granulocyte (neutrophil) antibodies can cause autoimmune neutropenia, drug-induced neutropenia, immune neutropenia after bone marrow transplantation, neonatal immune neutropenia, refractoriness to granulocyte transfusions as well as febrile and pulmonary transfusion reactions. In the last decade, considerable progress has been made in the characterization of the implicated antigens. In 1998, the Granulocyte Antigen Working Party of the ISBT introduced a new nomenclature for human neutrophil alloantigens (HNA), which is based on the antigens' glycoprotein location. In the HNA nomenclature the immunogenic (glyco-) proteins are indicated by arabic numbers followed by a letter of the alphabet which identify the (glyco-) proteins' polymorphisms, i.e. the specific antigens. Currently, seven HNA antigens are assigned to five systems. The HNA-1a, HNA-lb and HNA-1c antigens, the former NA1, NA2, and SH antigens, have been identified as polymorphic forms of the neutrophil Fc gamma receptor IIIb (CD16b) encoded by three alleles. Recently, we could elucidate the primary structure of the HNA-2a antigen, the former NB1. We could identify the HNA-2a-bearing glycoprotein as a novel member of the Ly-6/uPAR superfamily which has been clustered meanwhile as CD177. The HNA-3a antigen, the former 5b, is located on a 70-95 kDa glycoprotein. However, its molecular basis is still unknown. Finally, the HNA-4a and HNA-5a antigens, the former MART and OND, were found to be caused by single nucleotide mutations in the alphaM (CD11b) and alphaL (CD11a) subunits of the leucocyte adhesion molecules (beta2 integrins). The glycoproteins CD11b, CD16b, and CD177 have been found to be also frequent targets of autoantibodies - approximately 30% of neutrophil autoantibodies are directed against CD16b. Characterization of granulocyte antigens have expanded our diagnostic tools by the introduction of genotyping techniques and immunoassays for antibody identification. In addition, it allowed new insights in the pathophysiology of immune neutropenias and transfusion reactions. Ongoing studies will further improve the prevention and management of granulocyte antibody-mediated diseases.

Neutrophil alloantigens.

Antibodies to neutrophil antigens can cause neonatal alloimmune neutropenia, autoimmune neutropenia, febrile transfusion reactions, and transfusion-related acute lung injury. Several neutrophil antigen systems have been described serologically, but only the human neutrophil antigen-1 (HNA-1) or NA and HNA-2 or NB systems have been well characterized biochemically and molecularly. HNA-1 antigens are located on FcgammaRIIIb, CD16. HNA-2 antigens are located on 58- to 64-Kd glycoprotein, CD177, and are encoded by a gene on chromosome 19 that belongs to the Ly-6 family. The function of the CD177 is not known, but the CD177 gene is highly homologous to a gene overexpressed in neutrophils from patients with polycythemia rubra vera called PRV-1. New polymorphisms in these antigen systems are still being described, but the complete understanding of these neutrophil antigen systems has been slow because of the complexity of these genes.

Molecular nature of granulocyte antigens.

Granulocyte (neutrophil) antibodies can cause febrile and pulmonary (TRALI) transfusion reactions as well as immune neutropenias. In the last decade enormous efforts have been made to characterize the implicated alloantigens. The NA1, NA2, and SH antigens could be identified as polymorphic forms of the neutrophil Fc gammaReceptor IIIb encoded by three alleles. The antigens MART and OND have been located on the leucocyte adhesion molecules (beta2 integrins) and found to be caused by single nucleotide mutations in the alphaM (CD11b) and alphaL (CD11a) subunits. We have succeeded in throwing light on the primary structure of the NB1 antigen, which has recently been clustered as CD177. Based on these findings, the Granulocyte Antigen Working Party of the ISBT introduced in 1998 a new nomenclature for human neutrophil alloantigens (HNA nomenclature) based on the antigen's glycoprotein location. Elucidation of the molecular nature of granulocyte antigens now allows antigen identification by glycoprotein-specific immunoassays and genotyping by DNA techniques. Thus, considerable progress has been made in the characterization of granulocyte antigens. Further studies will improve our diagnostic tools and facilitate the prevention and management of transfusion reactions and immune neutropenias.

Molecular genetics of granulocyte polymorphisms.

BACKGROUND AND OBJECTIVES: Granulocyte (neutrophil) antibodies have been implicated in transfusion-related acute lung injury (TRALI) and immune neutropenias. In the last decade enormous efforts have been made to characterize the involved alloantigens. MATERIALS AND METHODS: Review of the literature. RESULTS: The NA1, NA2, and SH antigens have been identified as polymorphic forms of the neutrophil Fc gamma receptor IIIb encoded by three alleles. The antigens MART and OND have been located on leucocyte adhesion molecules (beta 2 integrins) and were found to be due to single nucleotide mutations in the alpha M (CD11b) and alpha L (CD11a) subunits. Recently, we succeeded in throwing light on the primary structure of the NB1 antigen. On the basis of these findings a new nomenclature has been introduced for human neutrophil alloantigens (HNA nomenclature). Glycoprotein location has made the development of antigen-specific immunoassays possible and antigen characterization on the molecular level now allows genotyping by DNA techniques. CONCLUSION: Considerable progress has been made in the characterization of granulocyte antigens. Further studies will improve our diagnostic tools and will facilitate the prevention and management of transfusion reactions and immune neutropenias.

Prognostic value of immunohistochemically detected CD44 expression in patients with carcinoma of the vulva.

BACKGROUND: Overexpression of alternatively spliced CD44 isoforms has been reported to correlate with poor prognosis in several human malignancies. To the authors' knowledge, there are no studies concerning the prognostic value of CD44 isoform overexpression in patients with squamous cell carcinoma of the vulva. METHODS: Thirty cases of squamous cell carcinoma of the vulva with International Federation of Gynecology and Obstetrics Stage I to III were examined immunohistochemically for overexpression of CD44 isoforms. We used 3 different variant exon sequence-specific murine monoclonal antibodies to CD44 isoforms containing variant exons v5, v6, and v7 to -8, respectively. The correlation of CD44 overexpression with clinical stage, histologic grade, and disease free and overall survival was investigated. RESULTS: CD44 isoforms CD44v5, CD44v6, and CD44v7-8 were detected in 83% (25/30), 63% (19/30), and 27% (8/30) of the tumor samples, respectively. Patients with tumors overexpressing CD44v6 showed a significantly shorter relapse free (log rank test, P = 0.002) and overall survival (log rank test, P = 0.003) compared with patients with tumors lacking CD44v6 overexpression. Expression of CD44v5 and CD44v7 to 8 had no impact on patients' survival. Clinical stage and histologic grade did not correlate with CD44 overexpression. CONCLUSIONS: Immunohistochemically detected overexpression of CD44 isoforms containing variant exon v6 is correlated with a poor relapse free and overall survival in patients with carcinoma of the vulva.

The 1990 Comprehensive Blood Bank Surveys of the College of American Pathologists.

The performance of participants on the 1990 Comprehensive Blood Bank Surveys of the College of American Pathologists was consistent with the high quality of past surveys. The surveys include challenges in ABO and Rh typing, crossmatching, transfusion decisions, unexpected antibody detection and identification, and supplemental questions to assess contemporary practices. As in previous years, a recurrent serologic problem has been the reporting of antibodies not present in serum (anti-E in Set J-B, and anti-K in Set J-D). Such overidentification of antibodies has little implication for transfusion safety and may, in fact, represent an artifact of the survey's samples and reporting practices.

[Detection of platelet-specific alloantigens in 400 unselected blood donors]. / Nachweis von plättchenspezifischen Alloantigenen bei 400 unausgewählten Blutspendern.

Antibodies against platelet specific alloantigens cause neonatal alloimmune thrombocytopenia, posttransfusion purpura, and they are sometimes found in polytransfused patients. In the last few years, new alloantigens were discovered in addition to the Zw-, Bak-, and Ko-alloantigens. In order to obtain representative data for the frequency of all platelet alloantigens in the European population, we typed 400 blood donors of our institution. No significant differences between our findings and data already published were found for the antigens of the HPA-1, -2, -3, and -5 systems; however, no HPA-4b (Yuka)-positive and no Naka-negative individual was found among the 400 blood donors tested. The Siba and HPA-2b antigens proved to be identical. The 'low-frequency' alloantigen Sra was not identified among the 400 individuals tested.

Expansion of the D system of horse red cell alloantigens.

Two additional specificities (Dq and Dr) were assigned to the D system of horse red cell alloantigens following discussion of the 1989 ISAG Horse Comparison Test (HCT) results. Family and population data support 25 phenogroups defined by the enhanced battery of 17 D system factors.

Report on the fourth ISBT/ICSH platelet serology workshop (London 1988).

The fourth ISBT/ICSH platelet serology workshop took place in 1988. It consisted of a wet workshop carried out by the 17 participating laboratories during March 1988, followed by a discussion of the results at a meeting of the ISBT/ICSH Expert Panel on Platelet Serology at the ISBT/BBTS Congress in London in July 1988. Sixteen samples, including 1 negative control serum, were analysed by the participants using methods of their choice. There was general agreement on 10/16 samples. The results indicated progress in the following areas: (i) antibody identification; (ii) differentiation of platelet-specific and HLA antibodies; (iii) glycoprotein localisation of platelet specific antigens; (iv) identification of new platelet antigen systems.

Expression of type 1 and type 2 blood group-related antigens in normal and neoplastic gastric mucosa.

The distribution of the blood group-related antigens type 1 (Lewis(a) [Le(a)], Lewis(b) [Le(b)]) and type 2 (H type 2, Y) has been examined in histologically normal and malignant mucosa of 40 surgical specimens of patients with adenocarcinoma of the stomach, with the use of a panel of monoclonal antibodies. Patients' Lewis phenotype and secretor status are correlated to the authors' findings. The surface epithelium of normal pyloric and fundic mucosa expressed the Lewis isoantigen (Le(a) in Le[a+b-] phenotype and Le(b) in Le[a-b+] phenotype), whereas the deep areas of this mucosa no showed the Le(a), Le(b) antigens and expressed the Y and H type 2 antigens whatever the secretor status of patients. Nineteen of 24 patients with Le(a-b+) phenotype showed anomalous expression of Lea antigen in neoplastic cells. In three of them, this alteration was found in tumor adjacent mucosa. No expression of Le(a) or Le(b) antigens was found in tumors or normal mucosa from Le(a-b-) phenotype patients.