Recognition by pregnancy serums of non-HL-A alloantigens selectively expressed on B lymphocytes.

A group of alloantibodies are found in pregnancy sera which react with antigens present on B lymphocytes and monocytes but are not detectable on the vast majority of unstimulated T cells. This specificity distinguishes them from HL-A antibodies which react with both cell types. They were readily recognized through indirect fluorescent antibody analysis by employing the combination of B-cell lymphoid lines and normal peripheral blood T cells. Different sera gave a variety of patterns of reactivity with a panel of 11 lymphoid lines. Similar differential patterns were also observed with normal B cells from different individuals particularly after concentrating the B cells. The antibodies were also cytotoxic to B cells and this procedure gave parallel results to the fluorescence method. The pattern of reactions obtained indicated a very heterogeneous system similar to that for HL-A. Special study of certain of the sera provided evidence that the lymphocyte-defined determinants of the mixed lymphocyte reaction system were involved. For convenience the term HL-B has been employed for these antigens.

Hereditary C2 deficiency: Genetic studies and association with the HL-A system.

Herediatary C2-deficiency has been shown to be transmitted asn an autosomal recessive characteristic. Recent evidence indicates that some genetic factors involved in the control of the complement (C) system in both man and mice are governed by genes localized within the major histocompatibility regionmthis study describes a large pedigree of the paternal family of a C2-deficient patient with systemic lupus erythematosusl It is shown that this condition is transmitted as an autosomal recessive trait, the heterozygous carriers having approximately half normal levels of C2. Furthermore, this trait was shown to be inherited in close linkage with an infrequent HL-A typw, 2,4A2. The maternal, C2-defective chromosome was shown to be transmitted by HL-AW10, W18 haplotypemthis same haplotype was described in a similar study by Fu et al. (6) to be associated with C2 deficiencymfinally, a third haplotype HL-A2,W18 carrying a defective C2 gene was demonstrated in a part of this pedigree.

Mixed lymphocyte culture determinants and C2 deficiency: LD-7a associated with C2 deficiency in four families.

Four families with C2 deficiency were studied. Among eight HL-A haplotypes involved with C2 deficiency, five were HL-A 10,W18. Three homozygotes for C2 deficiency from different families were mutually nonreactive in mixed lymphocyte cultures (MLC) and the heterozygotes from the fourth family failed to react to the homozygous cells. It appeared that identical MLC determinants were associated with all the genes from the different families that related to C2 deficiency. Further experiments identified the MLC determinant, LD-7a, as being involved. These results suggest marked linkage disequilibrium between the genes for C2 deficiency and the major histocompatibility complex (MHC). Studies of possible recombinants have offered tentative evidence for the positioning of the locus for C2 deficiency with respect to other segments of the MHC.

Cod liver oil inhibits neutrophil and monocyte chemotaxis in healthy males.

Epidemiological evidence suggests a reduced rate of chronic inflammatory diseases and ischaemic heart disease in populations with a high consumption of fish. This has been ascribed to the high content in sea food of polyunsaturated fatty acids (PUFAs), belonging to the n - 3 family. We have studied neutrophil and monocyte chemotaxis in 12 healthy males before and after 6 weeks supplementation with cod liver oil, corresponding to 5.3 g n - 3 PUFAs daily. Neutrophil and monocyte chemotaxis were investigated using the under agarose technique with N-formyl-methionyl-leucyl-phenylalanine (N-FMLP) and autologous serum as chemoattractants. Neutrophil chemotaxis towards both chemoattractants and monocyte chemotaxis towards N-FMLP were significantly reduced after supplementation with cod liver oil.

Real-time scanning and image analysis. A fast method for the determination of neutrophil orientation under agarose.

We describe a newly developed method for fast determination of neutrophil chemotaxis and orientation in concentration gradients of chemotactic factors. The system implements video-based real-time scanning and image analysis of neutrophil migration under agarose, using an interactive easy-to-use computer program. Two methods for determining cell orientation are presented. No statistically significant difference between the methods was found. The analysis program distinguishes between chemokinetic and chemotactic behaviour of the cells (P less than 0.01).

Detection of structural gene mutations and promoter polymorphisms in the mannan-binding lectin (MBL) gene by polymerase chain reaction with sequence-specific primers.

This study describes a new approach to the determination of all known mannan-binding lectin (MBL) mutations. The distribution of known variants of the MBL gene in a population of healthy unrelated Danes was determined and the genotype was correlated with the plasma MBL concentrations. The following genetic polymorphisms were studied: three point mutations in the promoter region at position -550 (H/L variants), -221 (X/Y variants), -70 (nt C or T), one point mutation in the 5' untranslated (UT) region at position +4 (P/Q variants) and three point mutations located at codons 52, 54 and 57 in exon 1 of the MBL gene, at nucleotide positions 223, 230 and 239, respectively. To perform genotyping, we designed sequence specific primers for a polymerase chain reaction (PCR-SSP). PCR-SSP is a powerful technique for the discrimination of alleles resulting from single base substitutions and is a widely used technique. Another major advantage of the PCR-SSP method is its ability to determine whether sequence motifs are in cis or trans. The frequencies of variants in exon 1 obtained by PCR-SSP were completely comparable to results obtained by previously described PCR methods, restriction fragment length polymorphism (RFLP) and site-directed mutagenesis (SDM). This PCR-SSP method is performed with standard laboratory equipment and has the capacity to detect all genetic variants in 100 samples in 2 days at an estimated total cost of GBP 11 per sample. Analysing the correlation between MBL haplotype and plasma MBL levels, we confirmed that three different structural variants, B, C and D and the promoter haplotypes HY, LY and LX have a dominant effect on the concentration of MBL. The HY haplotype is associated with the highest plasma concentration, the LY haplotype with intermediate levels and the LX haplotype with the lowest levels. The LX haplotype was found to be associated with very low levels of MBL similar to those found in association with the structural B genotype. The gene frequencies of variants in the MBL gene in the Danish population studied correspond to previous reports on Caucasian populations.

Effect of physical exercise on cytokines and lymphocyte subpopulations in human peripheral blood.

To examine the effect of intensive physical exercise on interleukin 2 (IL-2), tumor necrosis factor alpha (TNF alpha) and lymphocyte subsets, eleven elite and well-conditioned runners were tested in relation to a five-kilometer race. IL-2 was significantly decreased (p less than 0.01) immediately after the exercise and significantly increased after 24 hours (p less than 0.05), compared to the pre-exercise values taken at steady state. TNF alpha was significantly increased after 2 hours (p less than 0.05), and returned to habitual values after 24 hours. In the steady state at rest, elevation of HLA-DR+ cells was observed in all runners compared with control subjects (p less than 0.05), indicating a persistent activation of lymphoid cells. In connection with exercise a significant increase in NK cells (CD16+) was observed (p less than 0.01). The T-helper/T-suppressor (CD4+/CD8+) ratio was significantly reduced in connection with physical activity (p less than 0.01). In seven runners the ratio was reduced to a value of less than one. This decrease was observed immediately after the exercise, followed by increased ratios 2 hours later (p less than 0.01), due to oppositely directed quantitative changes of the CD4+ and CD8+ cell populations. After 24 hours the ratios returned to habitual levels. Furthermore, we confirmed an increase in the total number of granulocytes in connection with exercise (p less than 0.01), and observed a decrease in absolute numbers of lymphocytes two hours after exercise (p less than 0.01). We emphasize the importance of obtaining information about physical activity within the previous 24 hours before measuring white blood cell parameters.

HLA-C and -Bw typing of couples with unexplained recurrent miscarriages.

There is now evidence that the classical HLA class I molecule HLA-C is expressed on extravillous trophoblasts together with non-classical HLA-G molecules. Since clones of NK-cells are inhibited differently by supertypic epitopes associated with HLA-C and -B alleles we found it of interest to study HLA-C and -Bw polymorphism in 35 couples with recurrent miscarriage and 30 control couples with normal fecundity. All HLA assignments were undertaken by DNA techniques. The distribution of HLA-C alleles or the HLA-C associated supertypic epitopes recognized by NK1 or NK2 clones was not significantly different between patients and controls. The distribution of couples according to the number of NK1 and NK2 epitopes in the couple was similar in patients and controls. With respect to the HLA-Bw epitopes recognized by NKB1 clones, in 46% of the couples with recurrent miscarriage none of the spouses carried the HLA-Bw4 epitope compared with only 17% of the control couples (P < 0.02). It is concluded that the HLA-Bw4 epitope is carried more frequently by couples with normal fecundity than couples with recurrent miscarriage. The fetuses of couples with recurrent miscarriage are thus expected to lack expression of HLA-Bw4 epitopes on the trophoblast more often than fetuses of normal couples which might be of importance for the inhibition of NK-cell mediated antitrophoblast cytotoxicity.

HLA (SD and LD) in patients with amyotrophic lateral sclerosis (ALS).

Twenty-five patients with a definite diagnosis of amyotrophic lateral sclerosis (ALS) were HLA-typed for the serologically detectable antigens A, B and C and MLC-typed for 7 HLA-D-determinants. No significant deviation was found neither in the A, B, and C-series nor in the HLA-D-series as compared to normal controls. The aetiological problem of ALS is discussed.

Effect of human leukocyte interferon on human lymphocytes in vitro: cytogenetic studies.

Mitogen-stimulated lymphocytes from 8 healthy donors were exposed to interferon, and cytogenetic studies were preformed. The response of lymphocytes to the mitogens phytohemagglutinin (PHA), concanavalin A (con A) and pokeweed mitogen (PWM) was inhibited by interferon, whereas an increased number of structural chromosomal aberrations was not detected. Further investigations of the cytogenetic effects of interferon are needed.

Adult respiratory distress-like syndrome during hemodialysis: relationship between activation of complement, leukopenia, and release of granulocyte elastase.

Twelve patients with terminal uremia (8 females and 4 males) treated with chronic maintenance hemodialysis, were extensively studied during two successive dialyses with alternate use of either a Cuprophan (CP) based membrane, or a Polycarbonate (PC) membrane. Arterial plasma levels of total hemolytic complement, complement factors C3d and C5a, and granulocyte derived elastase were determined immediately before dialysis and sequentially during the entire procedure. Effluent line from the hemodialyzer was similarly sampled. Collected samples were centrifuged immediately at the bedside and instantly frozen in liquid nitrogen in order to preserve labile plasma components of complement. Analysis of the overall results shows that initial arterial leukopenia and generation of C5a in the hemodialyzer, as well as maximal values of hemodialysis-induced free plasma C3d and granulocyte elastase are related. Reflecting differences in biocompatibility, CP membranes were shown to induce significantly more leukopenia, increase in plasma free C3d, generation of C5a, and release of granulocyte-derived elastase. These results indicate that activation of complement, leukopenia, and release of granulocyte derived elastase are interlinked pathophysiological mechanisms of importance for acute deterioration of pulmonary function during hemodialysis, and that this condition is closely related to adult respiratory distress syndrome (ARDS).

[Lipids in the plasma of voluntary blood donors. A 2-year retrospective study (1985-1987)]. / Fedt i plasma hos frivillige bloddonorer. En 2 års retrospektiv undersøgelse (1985-1987).

In a regional blood transfusion centre, we observed 45 episodes of hyperlipidemic plasma among the 45,000 units produced from 12,000 unpaid volunteer blood donors during a two year period. Analysis of fasting serum cholesterol and serum triglycerides showed that 26 of the 45 were raised in either one or both of the values. Dietary instructions were given, in most cases through their general practitioners, who were informed of the results. Control of serum lipids taken in average 9.5 weeks later by the general practitioner showed a remarkable decrease in triglyceride and a slight decrease in serum cholesterol. Among the 26 we found one case of non insulin dependent diabetes mellitus, three possible and three chronic alcohol abusers. Based on these findings, fasting serum cholesterol and serum triglyceride are now analysed in donors, who repeatedly show hyperlipemic plasma.