Developing high-throughput HIV incidence assay with pyrosequencing platform.

Unlabelled: Human immunodeficiency virus (HIV) incidence is an important measure for monitoring the epidemic and evaluating the efficacy of intervention and prevention trials. This study developed a high-throughput, single-measure incidence assay by implementing a pyrosequencing platform. We devised a signal-masking bioinformatics pipeline, which yielded a process error rate of 5.

Comparing alloimmunization in preterm infants after transfusion of fresh unmodified versus stored leukocyte-reduced red blood cells.

Purpose: To compare the occurrence of red blood cell (RBC), platelet (PLT), and white blood cell (WBC) antibodies in preterm infants after transfusions.

Methods: A randomized, blinded trial was conducted in which preterm infants were transfused either with stored RBCs, prepared by prestorage leukocyte reduction and transfused throughout 42 days of storage to limit donor exposure (n = 18), or with fresh RBCs prepared without leukocyte reduction and transfused within 7 days after collection from as many donors as needed to guarantee freshness (n = 17). Nontransfused preterm infants of comparable birth weight were control subjects (n = 11).

Diversity associated with the second expressed HLA-DRB locus in the human population.

Although diversity within the HLA-DRB region is predominantly focused in the DRB1 gene, the second expressed DRB loci, DRB3, DRB4, and DRB5, also exhibit variation. Within DRB1(*)15 or DRB1(*)16 haplotypes, four new variants were identified: 1) two new DRB5 alleles, DRB5*0104 and DRB5*0204, 2) a haplotype carrying a DRB1(*)15 or *16 allele without the usual accompanying DRB5 allele, and 3) a haplotype carrying a DRB5(*)0101 allele without a DRB1(*)15 or *16 allele. The evolutionary origins of these haplotypes were postulated based on their associations with the DRB6 pseudogene.

Reconstruction of the immune system after unrelated or partially matched T-cell-depleted bone marrow transplantation in children: functional analyses of lymphocytes and correlation with immunophenotypic recovery following transplantation.

Reconstitution of the immune system following T-cell-depleted bone marrow transplantation (BMT) in children has yet to be fully elucidated. Thus, we prospectively studied the recovery of immune function in 64 children who underwent T-lymphocyte-depleted marrow transplants using either matched family member donors or matched unrelated donors. We measured in vitro posttransplantation proliferative responses to phytohemagglutinin (PHA), concanavalin A, pokeweed mitogen, and Candida albicans antigen and assessed unidirectional allogeneic mixed-lymphocyte culture (MLC) responses at various times.

A description of a new DR allele, DRB1*1113.

We have discovered a previously unpublished HLA-DRB1 allele, observed in a patient (SB), his mother, and one sibling. The undefined allele gave sporadic positive reactions with sera in the DR52-associated group. SSOPH analysis utilizing both generic and group specific primers and probes also gave ambiguous results.

Bw4-reactive and Bw6-reactive antibodies recognize multiple distinct HLA structures that partially overlap in the alpha-1 helix.

Bw4 and BW6 epitopes are expressed by mutually exclusive sets of HLA-B alleles and some HLA-A and HLA-C alleles. To test whether antithetical structures are required to express Bw4 and Bw6 epitopes, we measured binding of Bw4-reactive and Bw6-reactive alloantibodies and mAbs to HLA-B7 variants. A triple substitution of HLA-B7 alpha-1 helix residues 80, 82, and 83 created Bw4 and destroyed Bw6 epitopes detected by alloantibodies and mAbs.

The association of HLA-B8, B51, DR2, and multiple sclerosis in pars planitis.

Purpose: To establish a human leukocyte antigen (HLA) association in a homogeneous population of patients with pars planitis.

Methods: A strict set of inclusion parameters was established for the diagnosis of pars planitis. Forty patients with pars planitis who met these criteria underwent HLA analysis of class I and II phenotypes.

The relationship between HLA-A, B, DQ, and DR antigens and asbestos-induced lung disease.

To evaluate the relationship between human leukocyte antigen (HLA) and both asbestos-induced pulmonary fibrosis and pleural fibrosis, we obtained HLA-A, B, C, DQ, and DR phenotypes in 42 long-term asbestos-exposed workers. Among these exposed workers, 15 had asbestosis (ILO > or = 1/0) on the chest radiograph and 18 had asbestos-induced pleural fibrosis. We found that there was an increased percentage of HLA-A29, HLA-B44, and HLA-Bw4 in the subjects with asbestosis.

Salsalate-induced pancytopenia in a patient with systemic lupus erythematosus.

After a patient with a history of systemic lupus erythematosus in remission was given salsalate, all of her hematologic elements, especially leukocytes, profoundly decreased within hours, although the patient was receiving steroids. She had been challenged with salsalate at an earlier date, with a similar but less impressive drop in blood counts. Granulocytotoxic antibodies persisted during this episode in contrast to declining lymphocytotoxic and anti-native DNA antibodies that accompanied a remission of systemic lupus.

Critical role of HLA-DR beta 1 residue 58 in multiple polymorphic epitopes recognized by xenogeneic and allogeneic antibodies.

In a previous study, we identified glutamic acid at position 58 in DR (beta 1*1101) as critical for the epitopes recognized by the DRw11-specific mAb GS88.2, as well as the I-LR1 mAb that recognizes a polymorphic epitope on DR(alpha,beta 1*1101) and some DP molecules. The purpose of this study was to determine whether other polymorphic residues contribute to these epitopes and whether DR beta glutamic acid or alanine 58 and DP beta glutamic acid 56, the analogous position in DP beta, contribute to epitopes recognized by other anti-class-II mAb and allosera.

Biological mothers may be dangerous blood donors for their neonates.

Premature neonates require blood transfusions, and biological parents may wish to be directed donors. Biological mothers pose a potential danger because their plasma may contain antibodies that will react with blood cell antigens inherited by the infant from the father. We studied 25 healthy, pregnant women at the time of delivery for the presence of antibodies against red blood cell, leukocyte and platelet antigens.

HLA-B7 and HLA-DR2 antigens and acute posterior multifocal placoid pigment epitheliopathy.

Acute posterior multifocal placoid pigment epitheliopathy is a chorioretinal inflammatory disease occurring in young, healthy adults. Its cause is unknown, although it frequently follows a flulike illness. We reexamined 30 patients with documented acute posterior multifocal placoid pigment epitheliopathy to determine their HLA class I antigen (A and B) and class II antigen (DR and DQ) distribution.

Monocyte suppressor factor is plasminogen activator inhibitor inhibition of membrane bound but not soluble IL-1.

Human monocytes cultured in monolayer for 6 days were found to secrete a factor that suppressed the T cell proliferative response to soluble Ag and to alloantigens. The elaboration of this monocyte suppressor factor (MSF) was not inhibited by indomethacin. It has an apparent Mr of 50 to 60 kDa.

Low rejection rate when using unrelated or haploidentical donors for children with leukemia undergoing marrow transplantation.

Thirty-nine children with leukemia were entered into a bone marrow transplantation study comparing the use of unrelated donors who were HLA-Dr identical and matched at least with three of the four HLA-A or B loci, with haploidentical related donors. Although marrows were prepared by T lymphocyte depletion in vitro, the rejection rate of marrow in these patients was only 13% (5/39). Four patients had early rejections, and three of these four had autologous recovery immediately without evidence of leukemic recurrence.

Allogeneic marrow grafting with partially mismatched, unrelated marrow donors.

Forty patients with advanced hematologic malignancies or severe aplastic anemia received marrow grafts from partially mismatched, unrelated marrow donors. All patients were administered conventional prophylaxis for acute graft-v-host disease (GVHD) consisting of methotrexate and low-dose glucocorticoids. All but two patients who survived at least 30 days showed durable engraftment.

A near-fatal reaction during granulocyte transfusion of a neonate.

Although reactions to granulocyte transfusions in neonates are rarely reported, we observed a near-fatal pulmonary reaction, presumably due to white cell antibodies, in a neonate with Rh hemolytic disease. The hemolytic disease was being treated with exchange transfusions, and at 2 days after the infant's birth, bacterial sepsis was suspected and granulocyte transfusions were begun. The first granulocyte transfusion (Day 3) was uneventful.

Is secreted IL-1 necessary for normal human T-cell activation?

The role of secreted and membrane-bound IL-1 in the activation of human T cells by monocyte-associated, processed antigen was examined. The IL-1 is secreted from antigen-pulsed plastic-adherent monocytes for only 24 h after isolation. After extensive washing, however, these monocytes are fully capable of stimulating T cells to proliferate.

The effect of first cadaver renal transplant HLA-A, B match on sensitization levels and retransplant rates following graft failure.

Data were collected retrospectively on all 449 first-transplant cadaver renal allograft recipients transplanted at four centers between 1/1/78 an 12/31/82 who had graft failure by 1/1/85. A total of 383 of these patients had information available regarding subsequent disposition. Of these, 182 (47.

Resistance to suppression and acquisition of memory by human T cells.

Human memory cells acquire resistance to several types of suppressor cells, including MLR generated suppressor cells. These data suggest one possible mechanism of that resistance, namely retention of IL-2 receptors in the resting state. Cells from a 7-day MLR were separated on a single step percoll gradient.