Polymorphism of the complement receptor for C3bi.

RM2.184, a mouse IgG2a monoclonal antibody, recognizes a polymorphic determinant on the complement receptor for C3bi which is present on granulocytes and monocytes. The RM2.184 epitope is distinct from the monomorphic determinant recognized by the monoclonal antibody OKM1. The RM2.184 epitope is probably on the alpha subunit and dependent on the association of the alpha and beta subunits for its configuration, as it can not be detected after the subunits have been dissociated. The phenotypic frequency of the RM2.184 antigen is approximately 14%, and its segregation in families is independent of HLA and consistent with an autosomal co-dominant mode of inheritance.

Lymphscintigraphic evaluation of manual lymphatic drainage for lower extremity lymphedema.

To evaluate the effect of manual lymphatic drainage on technetium-99m-labeled dextran (99mTcDx) transport, 16 patients with lymphedema of lower extremities underwent two lymphscintigraphy exams by injecting 99mTcDx intradermally into the first interdigital space of the affected extremity. The first was a control examination at rest followed by an examination which included a manual lymphatic drainage session after the injection of the 99mTcDx. Images were obtained 45 minutes and three hours after the injection of the radioisotope. Extremity volumes were also measured before and after the drainage session. The findings from the examinations were assessed in a quantitative, semiquantitative and qualitative manner and compared without and with drainage. The analyses of the extremities' circumference before and after the drainage by paired t-test revealed a significant decrease. The analyses of the quantitative, semi-quantitative and qualitative evaluations evidenced no significant difference, without or with drainage, within the 45-minute and three-hour periods. Thus, manual lymphatic drainage caused an effective reduction in the circumference of the extremities but did not have a significant effect in the transport of 99mTcDx.

HLA-DR2 negative narcolepsy in Australian Caucasians: clinical features, serology and sequence specific oligonucleotide typing.

An almost invariable association with HLA-DR2 and DQw1 has previously been reported in Japanese and caucasian narcoleptics. We performed HLA typing in 18 Australian narcoleptics using serological techniques and sequence specific oligonucleotide probes. HLA-DQw1 was present in 15 patients and DR2 in 12; 3 patients with cataplectic narcolepsy were DR2-negative. The serological haplotype most strongly associated with narcolepsy was DRw15 (a subtype of DR2), DQw1. DRw15-positive patients were positive for the alleles DRB1*1501 and DQB1*0602 defined with oligonucleotide probes. We conclude that the association of narcolepsy with DR2 and DQw1 is not as strong as previously reported and the absence of DR2 or DQw1 does not preclude the diagnosis of classical narcolepsy, at least in caucasians. Secondly, DR2-positive narcoleptics possess characteristic serological subtypes and alleles defined with oligonucleotide probes that are also found in normals. Thirdly, the occurrence of DR2-negative cataplectic narcoleptics points to the existence of more than one narcolepsy susceptibility gene.

The short tandem repeat locus VWF2 in Intron 40 of the von Willebrand factor gene consists of two polymorphic sub-loci.

This study describes the complex nucleotide sequence structure of the TCTA short tandem repeat (STR) locus, VWF2. Eight alleles of VWF2 were observed in a population of 116 unrelated Caucasian individuals. The alleles ranged in size from 150 to 178 base pairs (bp). Sequence analysis of the isolated alleles revealed two polymorphic regions that were named sub-loci VWF2-a and VWF2-b. VWF2-a is located at the 5' end of the conventional locus, whilst VWF2-b is located at the 3' end. The two sub-loci are joined by a 30-nucleotide non-polymorphic sequence which contains two additional TCTA motif repeats. A semi-nested polymerase chain reaction (PCR) was designed to amplify the VWF2-b region in conjunction with the standard VWF2 amplification. This new amplification method enabled a higher level of allele discrimination than could be achieved by assigning alleles according to size. A cohort of 99 unrelated individuals was tested with this method. VWF2-a expressed five different alleles ranging from zero motif repeats to four motif repeats, while VWF2-b alleles ranged from 8 to 14 motif repeats. Allelic configuration based on the VWF2-a and VWF2-b sub-alleles revealed 23 unique configurations out of a possible 31 for the original eight VWF2 alleles. In conclusion, the VWF2 is a highly polymorphic STR locus with potential application for forensic and parentage testing.

ABO genotyping by polymerase chain reaction-restriction fragment length polymorphism.

Genotyping enables the identification of both maternally and paternally derived alleles. A number of protocols have been described for the genotyping of the ABO blood group system. Generally, these methods have a number of disadvantages including the use of hazardous reagents, being technically demanding, and the excessive use of materials. In this study, a relatively simple polymerase chain reaction -restriction fragment length polymorphism (PCR-RFLP) method is described. Four different amplifications were used that were specific for nucleotides sites 261, 526, 703, and 796 to distinguish the A, B, O1 and O2 alleles. The ABO genotypes of 294 random individuals were determined and were found to completely correlate with the serologic phenotypes. The protocol is applicable for investigations of weak or nonexpression of ABO alleles, paternity determinations, and population analysis.

ABO genotyping-identification of O1, O1*, and O2 alleles using the polymerase chain reaction-sequence specific oligonucleotide (PCR-SSO) technique.

ABO polymorphism at the gene level has been investigated by molecular methods, predominantly sequencing and restriction fragment length polymorphism (RFLP). We describe the application of the polymerase chain reaction-sequence specific oligonucleotide (PCRSSO) method, which is considered to be more versatile for large sample numbers, compared with conventional ABO genotyping by PCR-RFLP. PCR-SSO, while maintaining accurate and reliable results, reduces costs and labor. A population of 155 random individuals was investigated for the three O alleles, O1, O1*, and O2. The allelic frequencies were 35 percent, 26 percent, and 5 percent, respectively. PCR-SSO results correlated completely with both serologic and PCRRFLP results.

Serologic and molecular investigations of a chimera.

A chimeric individual possesses two or more genetically distinct cell populations. Although the chimerism may not be evident in all gene systems, various loci display greater numbers of alleles than genetically "normal" individuals. The proposita was referred for further laboratory investigation due to a mixed-field ABO blood group reaction following routine antenatal testing. Various molecular (HLA class II, ABO genotyping, and 10 short tandem repeat [STR] microsatellites) and serologic (HLA class I and red cell blood groups) typing techniques were employed to investigate a number of polymorphic loci located on different chromosomes. Chimerism was identified in 8 out of the 14 chromosomes tested: chromosome 1 (Duffy), 6 (HLA class I and II), 9 (ABO), 11 (HUMTH01), 12 (HUMPLA2A1), 15 (HUMFES/FPS), 18 (Kidd) and 21 (D21S11). The proposita was determined to be a probable dispermic chimera, based on the results of the serology and molecular studies.

Two novel alleles of the ATCT variable number tandem repeat locus VWF.VNTR I in intron 40 of the von Willebrand factor gene.

Two new alleles, allele 9 and allele 15, of the ATCT variable number tandem repeat locus, VWF.VNTR I, of intron 40 of the von Willebrand factor (VWF) gene are described. Both alleles occurred in low frequencies, being detected in only two and one of 285 random Australian Caucasians respectively. In all, 10 alleles were observed representing between six and 15 repeats of the ATCT motif. The allele frequency distribution in this Australian population was similar to other VWF.VNTR I population studies. The additional alleles described here for the VWF.VNTR I further enhance the usefulness of this VNTR locus in human identification work.

Instability in the ATCT variable number tandem repeat locus VWF.VNTR I in intron 40 of von Willebrand factor gene.

The von Willebrand factor gene intron 40 variable number tandem repeat VWF.VNTR I exhibits 10 alleles making it highly polymorphic and useful for parentage and forensic testing, 45 unrelated families (210 meiotic events) were tested for VWF.VNTR I alleles. One spontaneous mutation was observed in a family member. Haplotype analysis demonstrated that this mutation was due to a gain of one motif repeat by a paternal allele. Sequence analysis confirmed the difference in the number of motif repeats between the proband and the alleles expressed by the parents. This instability emphasizes the importance of demonstrating exclusion in at least two separate loci in parentage testing.

The IIb-IIIa glycoprotein complex that mediates platelet aggregation is directly implicated in leukocyte adhesion.

Evidence is presented that the IIb-IIIa glycoprotein complex, which functions as the receptor for fibrinogen on platelets and is central to platelet aggregation, is expressed on the surface of leukocytes where it may function as a receptor for fibronectin. F(ab')2 fragments of a monoclonal antibody, 25E11, raised against activated large granular lymphocytes, inhibited killing by natural killer cells, blocked the binding of fibronectin-coated particles by monocytes, and stimulated neutrophils to exhibit increased antibody-dependent killing. Immunoprecipitation studies of leukocytes and platelets, and the ability of 25E11 to inhibit platelet aggregation, identified the antigen as an epitope on the IIb-IIIa complex. This glycoprotein thus constitutes the first example of a receptor mediating both platelet aggregation and leukocyte adhesion.