William E. Kline

SEVERE PULMONARY HYPERSENSITIVITY ASSOCIATED WITH PASSIVE TRANSFUSION OF A NEUTROPHIL-SPECIFIC ANTIBODY☆

A life-threatening pulmonary hypersensitivity reaction accompanied by profound leucopenia and transient hypocomplementaemia developed in a 73-year-old man after repair of an abdominal aortic aneurysm. A neutrophil-specific antibody, anti-NA2) was demonstrated by granulocyte immunofluorescence and microagglutination in one of the transfused donor units. In addition, activated complement component C5a was generated in vitro by incubation of the patients serum with the aortic graft material. This severe clinical reaction may have resulted from the interaction of the transfused anti-NA2 antibody with complement activation induced by the aortic graft material.

Absence of HIV Infection in Blood Donors with Indeterminate Western Blot Tests for Antibody to HIV-1

To determine whether apparently healthy persons who have had repeatedly reactive enzyme immunoassays and an indeterminate Western blot assay for antibody to the human immunodeficiency virus type 1 (HIV-1) are infected with HIV-1 or HIV-2, we studied 99 such volunteer blood donors in a low-risk area of the country. The subjects were interviewed about HIV risk factors. Coded blood specimens were tested again for HIV-1 antibody (by two different enzyme immunoassays, a Western blot assay and a radioimmunoprecipitation assay) and for HIV-2 antibody by enzyme immunoassay, for HIV-1 by the serum antigen test, for HIV-1 by culture, for human T-cell leukemia virus Type I or II antibody by enzyme immunoassay, and for sequences of HIV DNA by the polymerase chain reaction. Of the 99 blood donors, 98 reported no risk factors for HIV-1 infection; 1 donor had used intravenous drugs. After a median of 14 months (range, 1 to 30) from the time of the initial test, 65 subjects (66 percent) were still repeatedly reactive for HIV-1 antibody on at least one immunoassay. In 91 subjects (92 percent) the Western blot results were still indeterminate, whereas in 8 they were negative. No donor met the criteria for a positive Western blot assay for HIV-1, and none had evidence of HIV-1 or HIV-2 infection on culture or by any other test. We conclude that persons at low risk for HIV infection who have persistent indeterminate HIV-1 Western blots are rarely if ever infected with HIV-1 or HIV-2.

Three Sera Defining a New Granulocyte ‐ Monocyte ‐T‐Lymphocyte Antigen

Abstract. Three sera containing antibodies recognizing a previously undescribed antigen on granulocytes were found during testing of sera from multiparous donors. All of the antibody producers were in good health. None had a history of transfusion. Using the granulocyte agglutination assay the sera recognize a single antigen which is not associated with the neutrophil antigens NA1, NA2, NB1, NC1, ND1, NE1, 5a, 5b, 9a, nor common red blood cell or HLA antigens. The three sera did not react with autologous cells or with the cells of the other antibody producers. Granulocytes from one antibody producer did not absorb antibody activity from any of the three sera. The antigen was found in large quantities on granulocytes and monocytes, in smaller quantities on T lymphocytes, and not on B lymphocytes, red cells, and platelets. The sera reacted with 340 of 343 random donors (99.1 %) and were negative with the same donor cells. Family studies showed autosomal dominant inheritance of the antigen. Five of 12 sibs in three families lacked this antigen (not statistically different from the expected ratio). The calculated gene frequency for the gene controlling the production of this antigen is 0.906. There appeared to be no association to the HLA, NA or Rh loci or to the X or Y chromosomes. None of the infants of these three women showed clinical signs of alloimmune neonatal neutropenia.

The frequency of granulocyte‐specific antibodies in postpartum sera and a family study of the 6B antigen

We examined sera from 2313 postpartum women as a potential source of granulocyte specific antibodies. Lymphocyte cytotoxic (LC) antibodies were detected in 397 (17.2%) specimens and granulocyte agglutinating (GA) antibodies in 291 (12.6%). Only two GA positive sera had reactivity for previously defined granulocyte specific antigens (one NA1 and NB1). One additional serum had GA reactivity similar to a serum previously reported by van Rood and called anti‐6B. The frequency of granulocyte specific antibodies (0.1 percent) in our study indicates that the productivity of random screening of parous sera for granulocyte specific antibodies is low. A four‐generation family study of the 6B antigen demonstrated a parallel association between HLA‐B7 and B40 and 6B reactivity in the LC, GA, and granulocyte immunofluorescence (GIF) assay. This illustrates that the granulocyte reactivity of anti‐6B is not granulocyte specific but is HLA related. This report provides additional evidence that HLA antibodies such as anti‐B7 (6B) can react with granulocytes bearing these antigens in the GA and GIF assays currently used to identify granulocyte specific antibodies.

Hepatitis B core antibody (anti-HBc) in blood donors in the United States: implications for surrogate testing programs

ABSTRACT: In order to evaluate the operational implications of excluding donated blood with antibody to hepatitis B core antigen (anti‐HBc), the American Red Cross tested 107,473 voluntary blood donations for anti‐ HBc in 72 test sites during a 1‐week period. The system‐wide prevalence of anti‐HBc was 2.60 percent, with a range of 0.55 to 6.38 percent, depending on geographic region. For the American Red Cross, which collects approximately one‐half of the blood supply in the United States, excluding donated blood with anti‐HBc would result in a loss of approximately 159,500 units during the first year.

Granulocyte Antigens and Antibodies

Granulocyte-specific antigens defined by human alloantisera are clinically important in neonatal neutropenia, autoimmune neutropenia, transfusion reactions, drug-induced immune neutropenia, and poor response to granulocyte transfusions. Many different antigens have been defined by human alloantisera using different assays. Only antigens of the N system of Lalezari are commonly studied today. The composition and location of these antigens within the granulocyte membrane are not known, but work is now proceeding on those issues. It also appears that these antigens may have some structural or functional role in the granulocyte. Although many myeloid MoAbs have been developed, there are very few reports to establish whether these identify structures that are related to the N series of granulocyte antigens. However, these MoAbs can serve as excellent reagents for better understanding the granulocyte membrane. It appears that studies during the next several years will provide exciting information about the structure-function relationships in the granulocyte membrane.

A microtiter modification of granulocyte immunofluorescence.

Abstract. The granulocyte immunofluorescence test (GIFT) is valuable for detecting allogeneic and autologous granulocyte antibodies. However, the original tube technique (macro GIFT) requires 50–100 μl of serum and 106 granulocytes with time‐consuming washing steps which cause cell loss. We report a modification (micro GIFT) using microtiter trays which requires only 20 μl of serum and 2 times 105 granulocytes. The modified method conserves reagents, reduces the time required for washing by 67%, decreases cell loss in washing by 74%, reduces the volume of conjugate required by 67%, and compares favorably with the macro GIFT in accuracy, specificity, and sensitivity.

Evaluation of clinical and laboratory aspects of antibody tests for detection of hepatitis C virus infection in blood donors and recipients from a low-risk population.

Background: When the first‐generation enzyme immunoassay (EIA) for detection of antibody to hepatitis C virus (anti‐HCV) was approved in May 1990, blood banking agencies recommended testing of all components in inventory. In many cases, one or more components from these units had already been transfused.

Autoimmune Neutropenia of Infancy and Early Childhood

Forty-one children were identified with autoimmune neutropenia of infancy and early childhood (absolute neutrophil count [ANC] less than 500/microliters and demonstrable serum antineutrophil antibodies). There were 21 boys and 20 girls; the median age at diagnosis was 11 months (range 5-38 months). No life-threatening infections occurred. There was a gradual upward trend in ANC in all patients over many months, with 87% having an ANC > 1000/microliters by 24 months from diagnosis. Among various clinical and laboratory parameters analyzed statistically, only younger age at diagnosis was associated with earlier neutrophil recovery. There was no association between degree or duration of neutropenia and sex, race, antibody reactivity, or presence of serious illness at diagnosis.

An Example of a Naturally Occurring Anti-cE (Rh27) that Binds Complement

A serum sample from a nontransfused male containing anti‐Rh27 and an additional weakly reacting antibody was investigated. One absorption of the serum with RzRl cells left only anti‐Rh27 while repeated absorption with RlRl cells or autologous cells had no effect. Treatment of the serum with reducing agents destroyed all activity suggesting the antibody was predominantly IgM. Tests with monospecific antiglobulin reagents in the indirect antiglobulin test revealed very weak reactions with anti‐IgM and anti‐IgA, no reactions with anti‐IgG, and strong reactions with anti‐C3(C3c + C3d) and anti‐C4. This is the first reported example of a naturally occurring complement binding anti‐Rh27.