Emily G. Reisner

The Complement-binding And Absorptive Capacity Of Human White Blood Cells Treated With Neuraminidase1

Human lymphocytes treated with Vibrio cholerae neuraminidase (VCN) are more susceptible to cytotoxic human alloantisera than are sham treated cells. Sera from most normal or immunized donors showing no detectable anti-HL-A activity gave strong positive reactions with almost all VCN-treated isologous or autologous lymphocytes. Less than 10% of the sera tested were unreactive against all VCN-treated cells. Family studies utilizing VCN-treated lymphocytes have revealed patterns of antigens that segregate independently from HL-A. The absorptive capacity of both VCN-treated and sham treated leukocytes for cytotoxic human alloantiserum is dependent on the class (reaction strength) of the ceEs used. Although VCN treatment does not affect the hierarchy of cell classes, which is based on HL-A reactions, VCN-treated cells absorb more antibody activity than sham treated cells. Absorption of a serum sample with sham treated leukocytes removed HL-A-related activity as detected by a panel of sham treated cells, but left antibody activity against VCN-treated cells. All detectable cytotoxic reactivity of sera against normal or VCN-treated lymphocytes was lost upon substitution of C6-defi-cient or cobra venom-treated rabbit serum for the routinely used complement source.SUMMARYHuman lymphocytes treated with Vibrio cholerae neuraminidase (VCN) are more susceptible to cytotoxic human alloantisera than are sham treated cells. Sera from most normal or immunized donors showing no detectable anti-HL-A activity gave strong positive reactions with almost all VCN-treated is

Paradoxical expression of adenosine deaminase in T cells cultured from a patient with adenosine deaminase deficiency and combine immunodeficiency.

T lymphocytes cultured from a patient (T.D.) with adenosine deaminase (ADA) deficiency expressed ADA activity in the normal range, inconsistent with her severe immunodeficiency, metabolic abnormalities, and with the absence of ADA activity in her B lymphocytes and other nucleated hematopoietic cells. ADA from T.D. T cells had normal Km, heat stability, and sensitivity to ADA inhibitors. Examination of HLA phenotype and polymorphic DNA loci indicated that T.D. was neither chimeric nor a genetic mosaic. Amplified and subcloned ADA cDNA from ADA+ T.D. T cells was shown by allele-specific oligonucleotide hybridization to possess the same mutations (Arg101----Trp, Arg211----His) previously found in the ADA-T.D. B cell line GM 2606 (Akeson, A. L., D. A. Wiginton, M. R. Dusing, J. C. States, and J. J. Hutton. 1988. J. Biol. Chem. 263:16291-16296). Our findings suggest that one of these mutant alleles can be expressed selectively in IL-2-dependent T cells as stable, active enzyme. Cultured T cells from other patients with the Arg211----His mutation did not express significant ADA activity, while some B cell lines from a patient with an Arg101----Gln mutation have been found to express normal ADA activity. We speculate that Arg101 may be at a site that determines degradation of ADA by a protease that is under negative control by IL-2 in T cells, and is variably expressed in B cells. Il-2 might increase ADA expression in T cells of patients who possess mutations of Arg101.

HLA-B15 association with erythema multiforme

Erythema multiforme (EM) is a cutaneous reaction pattern which follows numerous infections, drugs, neoplastic and inflammatory disorders in some individuals. We undertook a prospective study of thirty-eight HLA specificities of the -A, -B, and -C series in 16 Caucasian patients with EM and in 140 local Caucasian controls. Seven of 16 patients (44%) with EM and 5 of 9 patients (55%) with EM following herpes simplex infections possessed the HLA-B15 antigen, compared to 7% of local controls and 11.6% of the 1980 WHO Workshop Caucasian controls. Both associations were highly significant (p = 0.0125 and p = 0.02) when corrected for 38 HLA antigens. This is the first reported HLA association for erythema multiforme, a disease which may be a host-specific immune response to various antigens, determined in part by genes linked to HLA-B15.

Variable association of HLA-A2 in men with early-onset Alzheimer disease ☆

In the present study, we attempted to replicate the finding of an increased frequency of HLA-A2 in men with early-onset (less than or equal to 60 years) Alzheimer disease (AD). HLA data obtained on 167 patients (including 19 men with early-onset AD) from three geographic regions (North Carolina, Great Britain, and Finland) failed to replicate the result. A recent prospective study from Oregon, however, confirmed the association. Studies demonstrating the association suggest its presence in sporadic rather than familial AD. These results indicate a variable HLA/AD association, with some factor such as geographic region or disease familiality contributing to this variability.

A Feasibility Study of Human Leukocyte Antigen (HLA) Typing for Dried Bloodstains

This paper constitutes a feasibility report on the use of the human leukocyte antigen (HLA) system for the typing of dried bloodstains. Antigens tested include the HLA-A2, A3, A10, B7, B8, and B14 alleles. An aging study conducted on A3 positive bloodstains showed that HLA-A3 could be reliably detected on bloodstains stored up to 30 days at 22 degrees C. Unlike most earlier reports on HLA typing of bloodstains, no cross-reactivity problems were detected with the antisera used in this study. In addition to the successful typing of bloodstains, we were also able to type fresh, neat seminal and saliva stains in the A2 and A10 antigenic systems.

Application of probability of paternity calculations to an alleged incestuous relationship.

An alleged case of incest between half siblings has been examined by standard blood grouping and human leukocyte antigen (HLA) serology. The data were analyzed statistically using single and joint possibilities of paternity. The existence of the alleged relationship between the two parties in question is quite probable.

Tests of Genetic Markers on Aborted Fetal Material

Women who conceive as a result of rape often elect to abort the fetus. We describe twelve cases where genetic markers were tested on the aborted fetal material to provide evidence of the genetic constitution of the rapist. Two cases are presented in detail, and the problems encountered with the testing are discussed.

The Effect of Differences in Gene Frequency on Probability of Paternity

Knowledge of gene frequencies in populations is required for the calculation of probability of paternity. The question remains open as to the degree of accuracy of gene frequency estimates required to give accurate probability of paternity figures. This is of special concern in the HLA system, which has haplotype frequencies known to vary in populations. This paper presents computer simulation data comparing probability of paternity calculations using HLA data from California and North Carolina. Comparisons were made between geographic regions, and between blacks and whites within a geographic region. It was found that when the absolute probability of paternity is high, the average differences induced were small, but at lower probabilities the changes can be large. Differences were most pronounced between black and white populations. Examples of individual cases are given to illustrate the huge differences that can be induced in some cases by changing gene frequency.

54 ADENOSINE DEAMINASE (ADA) EXPRESSED AT HIGH LEVEL IN T LYMPHOCYTES OF A PATIENT WITH ADA DEFICIENCY AND SEVERE COMBINED IMMUNE DEFICIENCY DISEASE (SCID)

Among patients with ADA deficiency, those with SCID have near complete absence of ADA, while some patients with severe T cell dysfunction but preservation of B cell function may have 1-3% of normal ADA activity in lymphoid cells. Patients with >5% of normal ADA activity in lymphoid cells have had no immune defect. We describe a patient with classical onset of SCID at 3 months, who was begun on red cell transfusions and immunoglobulin replacement therapy at 6 months of age. Despite long term survival, she remained severely lymphopenic and immune deficient. After starting PEG-ADA therapy at age 10 years her lymphocytes increased and became responsive to mitogens, and she developed in vivo B cell function. Just prior to starting PEG-ADA, unfractionated nucleated cells from her bone marrow were found to have <1% of normal ADA activity, but IL-2 dependent T cells cultured from this marrow sample had near normal activity (>1000 nmol/h/mg protein). This observation was repeated with blood and marrow derived mononuclear cells and IL2-dependent T cells and T cell clones on 6 other occasions. HLA typing indicated the ADA+ T cells were the patients own cells. T cell ADA had normal heat stability. Km, pl and was inhibited >99% by EHNA. In contrast to her T cells, 12 EBV transformed B cell clones from the patient had <0.3% of normal ADA activity. ADA+ T cells had normal T cell surface antigens and responsed in vitro to IL-2, mitogens, antigen and allogeneic cells. Important questions raised by these findings concern the molecular basis for the T cell ADA activity and the reason that these cells failed to proliferate and function more effectively in this patient in vivo. Acquisition of ADA activity by T lymphocyte precursor cells may not be sufficient to restore immune function in some ADA deficient patients. This possibility should be considered in view of proposals to treat ADA deficiency by somatic cell gene replacement.