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Background BK polyomavirus (BKPyV) viremia/nephropathy and reduction in immunosuppression following viremia may increase the risk of alloimmune activation and allograft rejection. This study investigates the impact of BKPyV viremia on de novo donor anti-human leukocyte antigen (HLA)-specific antibodies (dnDSA). Patients and methods All primary renal transplants at East Carolina University from March 1999 to December 2010, with at least 1 post-transplant BKPyV viral load testing, were analyzed. Patients were negative for anti-HLA antibodies to donor antigens (tested via single antigen beads) at transplantation and at first BKPyV testing. Results Nineteen of 174 patients (11%) tested positive for BKPyV viremia. Within 24 months of BKPyV viremia detection, 79% of BKPyV-viremic patients developed dnDSA. Only 20% of BKPyV viremia-persistent cases, compared to 86% of BKPyV viremia-resolved cases, developed dnDSA (P = 0.03). Poor allograft survival was evident in BKPyV viremia-persistent patients (60% failure by 2 years post BKPyV diagnosis) and in BKPyV viremia-resolved patients with dnDSA (5-year post BKPyV diagnosis allograft survival of 48%). Conclusions Post-transplant BKPyV viremia and preemptive immunosuppression reduction is associated with high rates of dnDSA. When preemptively treating BKPyV viremia, dnDSA should be monitored to prevent allograft consequences.

The WHO Nomenclature Committee for Factors of the HLA System met following the 14th International HLA and Immunogenetics Workshop in Melbourne, Australia in December 2005 and Buzios, Brazil during the 15th International HLA and Immunogenetics Workshop in September 2008. This report documents the additions and revisions to the nomenclature of HLA specificities following the principles established in previous reports (1–18).

Lymphocytotoxic antibodies to the red cell blood groups A and B were purified from human sera by elution from synthetic immunoadsorbents (Synsorbs from Chembiomed). The lymphocytotoxic activity of the eluted antibodies was better preserved when elution was carried out at pH 11, whereas it was less stable when elution was performed at pH 3. In addition, complete removal of anti-red cell blood group activity from an anti-HLA serum was easily obtained by adsorption on the corresponding Synsorb without loss of anti-HLA activity.

T, B, and null lymphocytes isolated by sheep red blood cells (SRBC) rosetting or adherence to a nylon wool column had distinct size differences according to Coulter counter size analysis. The diameters in microns were as follows: B = 6.42, T = 6.76, null = 7.16, granulocytes = 8.42, and monocytes = 9.02. Coulter size distributions of cell preparations were compared to the proportions of cells with E, EAC, and surface immunoglobulin (SIg) markers in these preparations as well as their reactivity to HLA-DR antisera and rabbit anti-B-lymphocyte antisera. Cell preparations monitored by Coulter sizing produced the expected results in mixed lymphocyte culture (MLC) testing. That is, T cells responded but did not stimulate whereas B lymphocytes and monocytes stimulated but did not respond. NULL lympocytes both stimulated and responded. In antibody-dependent cell-mediated cytotoxicity (ADCC), null lymphocytes and monocytes functioned as effectors, whereas Tlymphocytes and granulocytes did not. B lymphocyte preparations were intermediate in activity. Coulter sizing is a rapid, simple means of assessing the purity of suspensions enriched for T, B, and null lymphocytes.

A hemolysis assay was developed to detect alloantibodies to human immunoglobulin. A total of 1035 serum samples was tested. Anti-IgM antibodies were found in 8% of 59 normal persons and in 13% of 439 multiparous women, with the highest incidence of 67% in 341 dialysis patients. Although the anti-IgM antibodies were inhibited by both IgM and IgG, it appeared that they were also inhibited by F(ab')2 but not by Fc. Anti-IgG antibodies were more strongly inhibited by Fc than F(ab')2. These results suggest that anti-IgM antibodies might be analogous to antiidiotypic antibodies directed to F(ab')2, whereas anti-IgG antibodies tend to have greater reactivity to Fc.

IgG and IgM were stripped from the surface of human B lymphocytes using heterologous anti-IgG or IgM antisera. The percent SIg (surface Ig) determined by fluorescence for SIgM was reduced from 55% to 20%, or a reduction of 64%. Upon stripping SIgM, the B lymphocytes were no longer killed by seven different cold cytotoxic antibodies obtained from kidney transplant patients. We conclude that SIgM is the target of cold cytotoxins. These cold cytotoxic antibodies are postulated to be immunoregulatory antibodies. They are less specific than anti-idiotypic antibodies.

To explore possible genetic determinants of immunity to Epstein-Barr (EB) virus infection, prevalence rates and geometric mean titers (GMT) of antibody against EB viral capsid antigen (VCA) were measured in 422 adults of known HLA and ABO histocompatibility types. Deviation from the overall prevalence (87%) of anti-VCA titers > 10 was limited to subjects with HLA-A10 (95%) and those with a "blank" HLA-A locus (78%). The latter deviation was largely accounted for by the 13 subjects of HLA-A1/blank constitution (46%). Deviations from the overall mean anti-VCA titer (GMT 131.5) were found in seropositive subjects with blood group 0 (GMT 153.3) and tissue antigens HLA-A3 (GMT 150.8), HLA-B7 (GMT 152.1), HA-Bw15 (GMT 97.0), and HLA-B27 (GMT 86.4). Separate reanalysis of the data for male and female subjects verified the association of blood group 0 with elevated and HLA-B27 with reduced mean levels of anti-VCA antibody.

The histocompatibility antigen, HL-A8, was found to be elevated in patients with Sjogren's syndrome. Among 24 patients, the frequency of HL-A8 was 58% compared to 21% among controls (P less than 0.001 after correction). With the addition of this disorder to the list of other autoimmune diseases associated with HL-A8, it now seems possible to postulate a single basis autoimmune defect gene in high linkage disequilibrium with HL-A8.

Plaque-forming cells can be generated from peripheral blood lymphocytes provided that the lymphocytes are prestimulated by in vitro sensitization in the presence of pokeweed mitogen. Of 91 blood type O persons whose lymphocytes were stimulated with red cell A antigen, 52% responded with plaque formation, whereas of 40 type O persons whose lymphocytes were stimulated with red cell B antigen, only 20% produced plaques. Similarly, reciprocal stimulation of type A and B persons showed higher plaque formation (65%) when 23 type B persons were stimulated with A than when 48 type A persons were stimulated with B red cells (19%). An optimal dilution of sensitizing cells is necessary and maximum time of sensitization was approximately 7 days.

The incidence of HL—A3 among 94 patients with a diagnosis of multiple sclerosis (MS) was significantly higher than for 871 normal persons. In addition, some increase in Te58 and decrease in HL—A2 and Te60 frequencies was noted. The well-known higher incidence of MS among Caucasians than among Orientals is paralleled by the higher incidence of HL—A3 and Te58 among Caucasians and a lower incidence of Te60.

Human lymphocytes sensitized with HL-A antibodies could not be killed by the addition of complement if the lymphocytes were incubated for periods of 1 to 5 hours at 37° C. This phenomenon was not produced by simple dissociation of the antibodies or by loss of antigens (modulation), but rather is postulated to result from an active release or pinocytosis of HL-A antigens together with the attached antibody. “Escape from sensitization” is followed closely by reformation of antigens, for the cells can readily be resensitized and killed by addition of complement. Loss of sensitization is an active process, since one-day-aged lymphocytes or lymphocytes treated with actinomycin D or puromycin were unable to express this activity. A differential escape rate for different specificities was encountered indicating that HL-A9 was produced more rapidly than several other specificities present on the same lymphocytes. A correlation was noted between the rate of HL-A antigen synthesis by lymphocytes (indicated by escape rate) and the quantity of HL-A antigen in the serum. Inhibition of lymphocyte cytotoxicity was used to detect the presence of HL-A antigens in serum. Certain specificities, such as HL-A9, were generally present in serum whereas others, such as HL-A8, could not be detected. Cross-inhibition tests with over 100 sera clearly showed an association of inhibition with specificity, although nonspecific inhibition was often observed. Sephadex fractions of sera tested at different concentrations revealed the greatest inhibitory activity in the 19S and 7S fractions.

Human peripheral blood granulocytes reacted with allogeneic or xenogeneic antisera and when stained with FITC formed polar caps very rapidly. Within 8 min, 90 % of the cells had clusters and polar caps. Cap formation generally paralleled the motility function of the cells. It occurred just as rapidly on granulocytes kept in hanging drops or in constant motion within a test tube as on glass slides. Phagocytic function of the cells was not affected by capping.

The matching criteria of counting the number of HL-A incompatibilities, the number of compatible antigens, and of ratios (net histocompatibility ratio) were evaluated. In order to avoid bias, the data were analyzed with the aid of computers, taking many different cutoff points. The probability value of statistical significance can be shown to vary considerably depending on the time selected for evaluating the clinical ranks and the clinical ranks which are considered to be “successful” or “failure.” Overall, the data indicate that because of the large number of patients who do well in spite of incompatibility, a more suitable means by which successful transplants can be predicted will be required. Since HL-A antigens are still the primary transplantation antigens (as shown by the successful HL-A-identical sibling transplants), empiric knowledge on the types of incompatibilities which can result in good graft survival must be developed.

Granulocyte cytotoxic activity in sera from over 257 patients was shown to be distinct from HL-A lymphocytoxic activity. Granulocyte cytotoxins occur in approximately 25 per cent of sera from patients having leukemia, 45 per cent with aplastic anemia, 22 per cent with kidney disease on hemodialysis, and 19 per cent of pregnant women. By testing sera on the same panel of cells, the granulocyte cytotoxic activity was shown not to be associated with granulocyte agglutination activity or lymphocytotoxic acitivty. It is likely that granulocyte cytotoxins and granulocyte agglutinins will be useful in transfusion and bone marrow transplantation as a separate tool from the more widely used lymphocyte cytotoxicity reaction.

We have defined the new allele HLA-B*4012, which had been isolated from a black individual. It was initially recognized as a serologically unique allele when typing her father for renal transplantation. The HLA class I phenotype was A*0201,*6602; B*4001,*4012; Bw6; Cw*0304,*1505. Sequencing from exon 1 through intron 3 of B*4012 was performed. B*4012 is identical to B*4001 and B*4010 in exon 3, and in the 3′ part of exon 2, but it is unique in that exon 1 and the 5′ part of exon 2 are identical to B*1503, B*1509, B*1510, B*1518, B*1523, and B*1529. The generation of this allele is best explained by a recombination event in exon 2 (break point between nucleotides 205 and 222 from the beginning of the coding region) of B*4001 or B*4010 with one of these B*15 variants as a donor allele. Its unique serological feature (B48, B60, B70, and B72 reactivity) is consistent with the sequence data of its donor alleles ( Note).

1. The Naming of New Genes Within the HLA Region The Committee agreed in 1996 to defer the naming of new genes within the HLA region. However, in view of the great interest and work being carried out on the MIC (MHC Class I chain related) genes, already named by the Human Genome Nomenclature Committee as MICA, MICB, MICC, MICD and MICE in April and September 1994 [15, 16], it was agreed that they should be included within the official HLA nomenclature for the purpose of naming alleles of these genes. In addition the change of name of the gene HLADNA to HLA-DOA, as published in the monthly updates, was confirmed, following further results showing that the gene codes for the alpha chain associated with HLA-DOB. The updated list of genes in the HLA region is given in Table 1.

We have characterized two novel HLA-B alleles, B*7802 and B*51022. The Caucasian-derived variant B*7802 most resembles the African-derived variant B*7801, from which B*7802 differs by two nucleotides. Only one of these modifications, however, is translated: a tyrosine for aspartate substitution occurs at residue 74 in B*7802, while the second nucleotide difference reflects a proximal synonymous substitution in codon 23. A second variant, B*51022, differs synonymously only at codon 23 from B*51021. Comparative analysis of the B5 CREG demonstrates that other pairs of B5 alleles differ synonymously only at codon 23 or synonymously at codon 23 and non-synonymously at a second more distal location. Contrary to the genesis of like pairs of B5 alleles via introduction of coordinate yet distant mutagenic events onto a single B5 progenitor, we postulate that synonymously different B5 progenitor molecules, B5ATT and B5ATC, are evolving in convergence to generate homologous B5 allele pairs differing silently at codon 23. Our finding that B*7802 is a single amino acid away from complete convergence with B*7801 and that B*51022 and B*51021 are in complete convergence is exemplary of such evolution.

We have developed monoclonal antibodies to most HLA specificities, making it possible for us to devise a simple, rapid, one-step microcytotoxicity test. The test is performed by adding 1 microliter of cells to 1 microliter of antibody-complement mixture predotted on the microtest tray. The reactions are read following a 1-hour incubation period (30 minutes in some instances). The analysis of reactions seen on testing 105 class I antibodies and 50 class II antibodies is shown. A comparison of typing by the standard NIH method and the new one-step procedure showed a > 96% concordance in the 500 T cells and 200 B cells we examined. Class I and class II typing could be performed using B cells, thus obviating the need to isolate both T and B cells for HLA typing.

HLA-B15 embraces a multiplicity of antigenic specificities which vary in their distribution amongst human populations. To correlate B15 molecular structure with the serological picture we have sequenced alleles encoding the various subspecificities of the B15 antigen: B62, B63, B75, B76 and B77, and a number of "variants" of these antigens including the 8w66 split of B63. HLA-B63 (B*1517) and 8w66 (B*1516) heavy chains have sequence identity to B17 in the alpha 1 helix correlating with the antigenic crossreactivity of these molecules. HLA-B77(B*1513) and B75 (B*1502) heavy chains differ solely in segments determining the Bw4 and Bw6 public epitopes, consistent with the serological description of the B77 and B75 antigens. One allele encoding the B76 antigen (B*1512) appears to be the product of gene conversion between the HLA-A and -B loci and differs from B*1501 in codons 166 and 167. In contrast, a second allele encoding the B76 antigen (B*1514) differs from B*1501 by an unrelated substitution in codon 167 which confers similarily with B45, an antigen crossreactive with B76. A third allele encoding B76, B*1519, differs from B*1512 by a unique point substitution in exon 4. Three alleles encoding variant B15 and B62 antigens (B*1508, B*1511 and B*1515) differ from B*1501 by localized clusters of substitutions that probably result from interallelic conversion. The B15 sequences described in this paper, in combination with those previously determined, define a family of 22 alleles, including those encoding the B46 and B70 antigens. Within this family the patterns of allelic substitution are analogous to those of other HLA-A and -B families, in that pairwise differences almost always involve functional positions of the antigen recognition site and recombination is the major agent of diversification.

The W H O Nomenclature Committee for factors of the HLA system met in Hakone after the 1 l th International Histocompatibility Workshop and Conference during November 1991 to consider additions and revisions to the nomenclature of specificities defined by both molecular and serological techniques following the principles established in previous reports [1-10] . The main subjects discussed were

Binding of immunoglobulins to major histocompatibility complex (MHC) molecules was demonstrated by two different assays: the binding of IgG to B cells by flow cytometry, and purified MHC antigens with an Elisa assay. Fc fragment from immune-complex binds to the Fc receptor on B lymphocytes. Here, Fab was also shown to bind to B cells. This binding was inhibited by specific human allo anti-HLA Class I and II sera directed at the polymorphic sites. Thus, in addition to the Fc receptor, MHC can also serve as a binding site for IgG. In an Elisa assay using purified antigens, IgG was shown to bind to HLA Class I and II molecules. Other proteins such as transferrin, human serum albumin, gelatin, etc., did not bind to the MHC proteins. Immunoglobulins bound to MHC molecules by sites on the Fab fragment independent of the hypervariable region. This was demonstrated by the retention of antibody activity even after binding of antibody (anti-lactoferrin) to MHC. The relative avidity between Fab and HLA Class I and II was 4-8 x 10(5) M-1.

Idiopathic generalized epilepsies, i.e., juvenile myoclonic epilepsy (JME), childhood absence epilepsy, and epilepsy with grand mal [generalized tonic-clonic seizures (GTCS)], are the most common genetic epilepsies. Linkage studies using Bf, HLA serologic, and DNA markers by three independent investigators, one from Los Angeles and two from Berlin, have localized the JME locus to the short arm of chromosome 6 (6p). Because members of the same JME family have the same JME phenotype of childhood absence epilepsy, epilepsy with grand mal (GTCS) seizures, or early childhood myoclonic epilepsy (ECME), our observations give evidence for a single-locus etiology in 6p for JME and for at least some of the childhood absence seizures, epilepsy with grand mal (GTCS) seizures, and ECME. Studies should now address whether locus heterogeneity exists within childhood absence epilepsy, epilepsy with grand mal (GTCS) seizures, or ECME. Markers linked to JME (Bf, HLA serologic, and DNA markers in the DQ region) can be used to resolve etiologic heterogeneity. Using such markers, both linked and unlinked forms of phenotypes that are clinically indistinguishable may be detected and provide evidence for etiologic heterogeneity. Studies should also concentrate on narrowing the JME locus to 2 to 3 cm by screening families with recombinant events using RFLPs, candidate genes, and new expressed sequences on chromosome 6.

To test the humoral theory of allograft rejection, we investigated whether all patients who reject a kidney have HLA-antibodies. Sera from 1363 patients at four transplant centers who had rejected a kidney graft and transplant or who had rejected a graft and were waiting for a regraft were first tested by cytotoxicity AHG. Sera that were negative were tested by solid phase assays (ELISA or flow cytometry). Among the 1363 patients who had rejected a kidney, 90% were found to have HLA-antibodies. Eighty-four per cent tested positive for the CDC-AHG assay and an additional 6% of sera, when tested by solid-phase assays for IgG, IgM and IgA antibodies, was found to be positive for HLA. Among the four different centers the range of positivity was 89–91%. We conclude that 90% of patients who reject a kidney transplant have detectable HLA-antibodies.

Nineteen patients with Sjögren's syndrome were evaluated for the presence of HLA-B8 and HLA-Dw3. HLA-B8 was found in 57.8% of patients and 20.1% of 96 controls (P less than 0.001). HLA-Dw3 was detected in 73.7% of the patients and 24.0% of controls (P less than 0.0001). Statistical evaluation of the association of both antigens with Sjögren's syndrome revealed that the primary association was with HLA-Dw3 (P less than 0.005). Patients with HLA-Dw3 had a lower mean parotid salivary flow rate (0.8 +/- 0.3 ml/10 minutes/gland versus 5.6 +/- 1.2 ml/10 minutes/gland, P less than 0.0004) and a more intense lymphocytic infiltration into labial salivary glands (focus score 7.2 +/- 0.9 versus 3.4 +/- 0.9, P less than 0.04) than did patients without HLA-Dw3. The difference in the intensity of the lymphocytic infiltration was even more significant when patients with both HLA-Dw3 and HLA-B8 were compared with patients with neither antigen (focus score 7.5 +/- 1.0 versus 2.8 +/- 0.8, P less than 0.02). In addition, all patients with severe keratoconjunctivitis sicca had both antigens. Our observations suggest that a number of genes may interact to determine susceptibility and severity in Sjögren's syndrome.

Immunologic approaches to the classification of acute lymphocytic leukemia (ALL) have led to a new awareness of the heterogeneity of this disease. Surface membrane markers including surface membrane immunoglobulin, complement receptors, and sheep erythrocyte (E) receptors have demonstrated at least three subtypes of ALL, which include non-B, non-T, ALL, T-ALL, and B-ALL. In addition, hetero-antisera to Ia-like antigens and ALL-associated antigens have been used to positively identify non-B, non-T ALL, which was previously a diagnosis of exclusion. This paper reports 17 cases of childhood ALL whose lymphoblasts were studied for surface membrane immunoglobulin, sheep erythrocyte receptors, and the presence of four antigens detected by well-characterized heteroantisera. Every non-B, non-T lymphoblast was positively identified by the Ia-like antiserum and/or the ALL antiserum. One T lymphoblast was identified by E receptors and the T antiserum, whereas two did not have E receptors but did react to the T antiserum. None of these three T lymphoblasts reacted with the Ia-like antiserum.

Of 39 consecutive cadaver donors tested, DR antigens could not be detected in 17. Following 3 h incubation of the B lymphocytes at 37 degrees C, eight of the donors with no detectable DR antigens had two DR antigens and nine had one DR antigen. In addition, there were nine cases in which a potential cadaver donor had one antigen before incubation and two DR antigens following incubation. The simple technique of pre-incubating B lymphocytes before testing has helped considerably in prospectively typing cadaver donors for HLA-DR.

HLA antigens were examined by microdroplet lymphocyte cytotoxicity in 31 patients with Sjogren's syndrome. Fifty percent of 22 Caucasian patients (7 of 17 females and 4 of 5 males) had HLA-B8, as compared to 21% of 1205 controls (P = 0.002). The presence of HLA-B8 was not correlated with parotid salivary flow, kerato-conjunctivitis sicca, or focus score. Because of its association with a number of autoimmune diseases, including Sjogren's syndrome, it is suggested that HLA-B8 is genetically linked to an immune response gene(s) that predisposes the individual to autoimmune phenomena. The expression of autoimmunity may be determined by infectious or environmental factors.

For the past 10 years, the International Cell Exchange has been a useful tool for testing rare and unusual HLA antigens and monitoring the progress in detection of recognized specificities. Of special interest are the antigens proposed in the 8th Workshop - 8w57, 8w58, 8w59, and 8w66. Marked improvement could be seen in the detection of 8w59 (29%) and 8w66 (31%). Possible new variants of the A9 and A10 complexes were tested in several exchanges. As shown by the increasing yearly average percentages of detection, laboratories are improving in their ability to type Aw23, Aw24, A25, A26, A29; Bw39, Bw41, Bw54, Bw55, Bw57, Bw58, Bw60 and Bw62. Over 90% of the laboratories are now able to detect Aw24; B5, B17, and B40, and A29 has attained the status of being well defined.

Highly enriched preparations of monocytes, B and T lymphocytes, and granulocytes from 18 normal donors were serotyped in parallel in a complement-dependent cytotoxicity assay using allogeneic and heterologous antisera defining three independent tissue antigen systems. HLA and B-lymphocyte tissue antigens were detected on human monocytes although granulocyte antigens were absent. By cytotoxicity testing the presence of Ia-like antigens on monocytes was significantly diminished compared to the autologous B-lymphocyte population and has important implications in B-lymphocyte serology. The study indentified a number of human antisera obtained from multitransfused subjects and pre- and post-transplant organ recipients that were non-HLA and appeared to define monocyte-associated antigens. The serological implications of surface antigen expression on human monocytes compared with other peripheral blood cells are discussed.

Spleen cells from a mouse immunized with an AML cell expressing HLA-A3 produced a hybridoma secreting an anti-HLA-A3, A11 monoclonal antibody, 26D3. By complement-dependent cytotoxicity at dilutions to 1:10(4) the ascites antibody lysed 13/13 A3, 15/15 A11, and 8 other lymphocytes from a panel of 98 donors. The 26 D3 immunoprecipitated a molecule consisting of subunits of 44,000 and 12,000 daltons. Monoclonality of the antibody was demonstrated by isoelectric focusing and protein A affinity chromatography.

Anti-Fab antibody titers were positive in 70% of rheumatoid arthritis patients (54 of 77) and 3% (1 of 35) of healthy donors. Their specificity was examined by inhibition of the radioimmunoassay for anti-Fab antibodies, which demonstrated that they are against the Fd region of the intact immunoglobulin molecule. In addition, anti-Fab antibodies have broad specificities for IgG antigens, shown by inhibition with myeloma IgG. They are cytotoxic against B (20% of B cells from most healthy donors) but not T lymphocytes, with cytotoxicity greater at 5 degrees C than 37 degrees C. These studies show that the anti-Fab antibodies interact with allotypic or idiotypic determinants on subpopulations of B lymphocyte cell surface antigens.

Actual HL-A typing has been performed on 115 members of cancer family N, a large kindred (over 1000 members ascertained) showing the findings consistent with the cancer family syndrome. In the cancer-prone line (branches C and D) of the family, 20 of 21 members with cancer had one HL-A haplotype, HL-A2-HL-A12 (relative odds = 6.30), including some decreased family members who had haplotypes assigned. Eleven of 12 family members with cancer in branches C and D, actually typed, had HL-A2-HL-A12 (relative odds = 6.06). The single exception showing cancer and another haplotype in branch D is a child of a family member with haplotype HL-A2-HL-A12.

The etiology of Hodgkin's disease is complex, as is evident in studies suggesting the importance of horizontal transmission, occupational factors, racial and ethnic background, and familial, genetic factors, or both, including HL-A associations. The present study is of a remarkable kindred in which Hodgkin's disease was histologically verified in two sibships involving second-cousins related through maternal great-grandparents. Cancer of the lung, breast, endometrium, ovary, pancreas, and brain, as well as leukemia and Wilms' tumor, occurred in first and second-degree relatives of the Hodgkin's patients. HL-A haplotypes in patients with Hodgkin's disease in this family showed HL-AB5 or HL-ABW35, an association confirmed in other reports. The findings of associated malignant neoplasms in familial Hodgkin's disease, here and elsewhere in the literature, suggest that in the quest for etiology of Hodgkin's disease one must view the disorder eclectically, with a painstaking search for multiple etiologies, genetic and environmental.

The sera of H-normal, H-weak and H-deficient individuals transferred the same amounts of ABH and Lewis antigens to lymphocytes in culture, confirming that the circulating ABH and Lewis antigens detected by lymphocytotoxicity are independent of the H-h system. These antigens were, as expected, under the control of the secretor and Lewis systems in the same way as exocrine secretions. These results suggest that both circulating ABH and Lewis glycosphingolipids and exocrine ABH and Lewis glycoproteins can be synthesized by the same tissues.

Comparison was made of HL-A phenotypes in 122 patients with systemic lupus erythematosus (SLE) and 1289 ethnically matched controls. The only significant association was between the HL-A antigen HL-A5 and the presence of SLE in Black Americans. A similar trend in 40 Caucasian patients did not reach a level of statistical significance. No significant association was present between any HL-A antigen and clinical or laboratory manifestations of the disease. Lymphocytotoxic antibodies were present in 36% of the patients and correlated with the presence of low total hemolytic complement and leukopenia.

Although certain forms of epilepsy have long been suspected to be inherited, heterogeneity has made it difficult to find the genes responsible for any subtypes. We found that families ascertained through patients with juvenile myoclonic epilepsy show linkage with the BF and HLA loci on human chromosome 6. There is some evidence that the locus may be outside the HLA complex and no evidence as yet of an association with any allele of the HLA complex.

SUMMARY Fifty-seven members from ten families in which one parent and at least one child have asthma were studies with dilutional skin tests and RAST to grass pollens after determination of HLA haplotypes. We found no direct evidence for linkage of a hypothetical asthma locus with HLA or for a significant association of asthma with HLA haplotypes. Linkage between the HLA loci and a gene or genes which allow for the expression of clinical asthma could neither be proven nor disproven due to the small sample size. All of the asthmatic children had positive dilutional skin tests and RAST, suggesting that atopic asthma may be genetically controlled by the HLA chromosome (chromosome 6). Nonetheless, determination of the histocompatibility antigens can increase the value of predictive risk analysis for asthma. Such a determination may be important in the early identification of a child born to a family with atopic asthma.

Two EBV-negative lymphoma lines of human B-cell origin, BJAB and Ramos, were compared with altogether six of their in vitro EBV-converted, EBNA- and EBV-DNA-carrying sublines (four of Ramos and two of BJAB derivation). All converted lines closely resembled the parental line with regard to karyotype and HL-A and B antigen typing. Induction of EBV antigens (EA and VCA) by P3HR-1 virus superinfection was either similar in the converted and the negative lines, or somewhat increased in certain converted lines. These findings argue against a simple, virally determined repressor model and emphasize the role of cellular controls in restricting the EBV cycle in virus-carrying B-lymphocyte lines of human origin. IUdR inducibility varied in the different converted lines. There was a possible relationship between average number of EBV-genome equivalents per cell and inducibility. Converted sublines did not differ from the original negative lines with regard to surface immunoglobulin and Fc receptors. There was a dramatic increase in complementconsuming ability, however, following EBV conversion. Among the EBV-positive lines, there was a linear relationship between complement-consuming and EBV-receptor activity, the latter measured by a quantitative absorption test. Inductibilite Du Cycle Du Virus D'Epstein-Barr (EBV) Et Proprietes Des Marqueurs De La Surface Des Lignees De Lymphome EBV-Negatives Et De Leurs Sous-Lignees Transformees in vitro par L'EBV Deux lignees de lymphome EBV-negatives provenant de cellules B humaines, BJAB et Ramos, ont ete comparees avec six de leurs sous-lignees transformees in vitro par l'EBV et portant l'EBNA et l'ADN de l'EBV (quatre sous-lignees Ramos et deux BJAB). Toutes les lignees transformees ressemblaient etroitement a la lignee parentale du point de vue du caryotype et du typage des antigenes B et HL-A. L'induction des antigenes de l'EBV (EA et VCA) par surinfection avec le virus P3HR-1 etait soit analogue dans les lignees transformees et les lignees negatives, soit legerement superieure dans certaines lignees transformees. Ces constatations ne concordent pas avec l'hypothese d'un modele de represseur simple, determine par le virus, et mettent en evidence le rǒle des contrǒles cellulaires dans la restriction du cycle de l'EBV dans les lignees de lymphocytes B d'origine humaine qui hebergent le virus. L'inductibilite sous l'effet de l'IUdR varie selon les lignees transformees. Il existe peut-ětre un rapport entre le nombre moyen d'equivalents du genome EBV par cellule et l'inductibilite. Les lignees transformees ne different pas des lignees negatives initiales du point de vue de l'immunoglobuline de la surface et des recepteurs Fc. On a toutefois constate une augmentation frappante de la capacite d'epuisement du complement apres la conversion par l'EBV. Dans les lignees EBV-positives, on a note un rapport lineaire entre l'epuisement du complement et l'activite des recepteurs de l'EBV, cette derniere ayant ete mesuree au moyen d'un test d'absorption quantitatif.

The variable expression of the C locus specificities at the cell surface is a major problem in HLA typing and the phenomenon was clearly documented by the International Cell Exchange Program. In contrast with HLA-A and -B, the C locus specificities are readily lost from cells following storage at room temperature for a few days or in overnight incubation at 37 degrees C in the presence of protein synthesis inhibitors. These specificities can be made to reappear by making the cells metabolize at 37 degrees C for 8 to 20 h in nutrient medium. Sixty-nine per cent of the positive C locus reactions lost after 7-day storage at room temperature were recovered after overnight incubation at 37 degrees C. The reappearance was dependent on protein synthesis since presence of cyclohexamide during incubation completely blocked the recovery. A simple overnight incubation is very useful in enhancing C locus expression on older cells.

The records of thirty-one patients with nasopharyngeal carcinoma (NPC) diagnosed from 1966 through 1976 among the Alaskan native population (Eskimo, Aleut, Indian) were reviewed. There were 25 males and six females, which results in relatively high incidence rates per 100,000 of 13.5 for males and 3.7 for females. Clinical and pathologic features were similar to those found among southern Chinese NPC patients. Five-year survival rate was 48%. Antibodies to Epstein-Barr virus were higher in NPC patients than in patients with other tumors or matched controls. On histocompatibility testing Sin-2 was not detected, nor was there significantly increased frequency of A2. Instead, BW40 and a second locus blank occurred more often among NPC patients than among other groups. In response to a questionnaire, NPC patients more often reported use of salt fish in the childhood diet, smoking of cigarettes, and exposure to noxious inhalants than did controls, but the differences were not statistically significant.