In most PCR-based tissue typing techniques the PCR amplification is followed by a post-amplification specificity step. In typing by PCR amplification with sequence-specific primers (PCR-SSP), typing specificity is part of the amplification step, which makes the technique almost as fast as serological tissue typing. In the present study primers were designed for DR "low-resolution" typing by PCR-SSP, i.e. identifying polymorphism corresponding to the serologically defined series DR1-DRw18. This resolution was achieved by performing 19 PCR reactions per individual, 17 for assigning DR1-DRw18 and 2 for the DRw52 and DRw53 superspecificities. Thirty cell lines and 121 individuals were typed by the DR "low-resolution" PCR-SSP technique, TaqI DRB-DQA-DQB RFLP analysis and serology. The concordance between PCR-SSP typing and RFLP analysis was 100%. The reproducibility was 100% in 40 samples typed on two separate occasions. No false-positive or false-negative typing results were obtained. All homozygous and heterozygous combinations of DR1-DRw18 could be distinguished. Amplification patterns segregated according to dominant Mendelian inheritance. DNA preparation, PCR amplification and post-amplification processing, including gel detection, documentation and interpretation, were performed in 2 hours. In conclusion, PCR-SSP is an accurate typing technique with high sensitivity, specificity and reproducibility. The method is rapid and inexpensive. DR "low-resolution" typing by the PCR-SSP technique is ideally suited for analyzing small numbers of samples simultaneously and is an alternative to serological DR typing in routine clinical practice including donor-recipient matching in cadaveric transplantations.

HLA-DR typing by PCR amplification with sequence-specific primers (PCR-SSP) in 2 hours: an alternative to serological DR typing in clinical practice including donor-recipient matching in cadaveric transplantation.

A large case-control study of malaria in West African children shows that a human leucocyte class I antigen (HLA-Bw53) and an HLA class II haplotype (DRB1*1302-DQB1*0501), common in West Africans but rare in other racial groups, are independently associated with protection from severe malaria. In this population they account for as great a reduction in disease incidence as the sickle-cell haemoglobin variant. These data support the hypothesis that the extraordinary polymorphism of major histocompatibility complex genes has evolved primarily through natural selection by infectious pathogens.

/pdf/common-west-african-hla-antigens-are-associated-with-22igsgshrl.pdf

Common West African HLA antigens are associated with protection from severe malaria

This report has been prepared by an EAACI task force representing the five EAACI Sections and the EAACI Executive Committee composed of specialists that reflect the broad opinion on allergy expressed by various clinical and basic specialties dealing with allergy. The aim of this report is to propose a revised nomenclature for allergic and related reactions that can be used independently of target organ or patient age group. The nomenclature is based on the present knowledge of the mechanisms which initiate and mediate allergic reactions. However, the intention has not been to revise the nomenclature of nonallergic hypersensitivity.

A revised nomenclature for allergy. An EAACI position statement from the EAACI nomenclature task force.

Molecular mimicry in T cell-mediated autoimmunity: Viral peptides activate human T cell clones specific for myelin basic protein

We report a natural history study of 216 patients with primary progressive (PP)- multiple sclerosis defined by at least 1 year of exacerbation-free progression at onset. This represents 19.8% of a largely population-based patient cohort having a mean longitudinal follow-up of 23 years. This subgroup of PP-multiple sclerosis patients had a mean age of onset of 38.5 years, with females predominating by a ratio of 1.3:1.0. The rate of deterioration from disease onset was substantially more rapid than for relapsing-remitting multiple sclerosis, with a median time to disability status score (DSS) 6 and DSS 8 of 8 and 18 years, respectively. Forty-nine percent of patients were followed through to death. Examination of the early disease course revealed two groups with adverse prognostic profiles. Firstly, a shorter time to reach DSS 3 from onset of PP-multiple sclerosis significantly adversely influenced time to DSS 8. Second, involvement of three or more neurological systems at onset resulted in a median time to DSS 10 of 13.5 years in contrast to PP-multiple sclerosis patients with one system involved at onset where median time to death from multiple sclerosis was 33.2 years. However, age, gender and type of neurological system involved at onset appeared to have little influence on prognosis. Life expectancy, cause of mortality and familial history profile were similar in PP-multiple sclerosis and non-PP-multiple sclerosis (all other multiple sclerosis patients from the total population). From clinical onset, rate of progression was faster in the PP-multiple sclerosis group than in the secondary progressive (SP)-multiple sclerosis group. When the rates of progression from onset of the progressive phase to DSS 6, 8 and 10 were compared, SP-multiple sclerosis had a more rapid progressive phase. A substantial minority (28%) of the PP-multiple sclerosis cohort had a distinct relapse even decades after onset of progressive deterioration. These studies establish natural history outcomes for the subgroup of multiple sclerosis patients with primary progressive disease.

The natural history of multiple sclerosis: a geographically based study. 5. The clinical features and natural history of primary progressive multiple sclerosis.

In tissue culture, immune lymph node cells containing foreign histocompatibility antigens of the H-2 type exert marked cytotoxic effects on tumor cells incompatible with the H-2 antigen. An equally pronounced effect is obtained when normal allogeneic and semi-isologous lymphoid cells of F 1 hybrids are caused to aggregate around the target tumor cells by treating the cultures with either heat-inactivated rabbit antiserum to mouse cells or phytohemagglutinin. Isologous lymph node cells have no effect. Thus, aggregation of lymphoid cells and target cells is a necessary but insufficient requirement for cytotoxicity in vitro; in addition, close contact must be established between histoincompatible cells.

Contact-induced cytotoxicity by lymphoid cells containing foreign isoantigens.

HLA class II gene polymorphism was investigated in 100 patients with clinically definite multiple sclerosis (MS) by restriction fragment length polymorphism analysis of Taq I-digested DNA using DRB, DQA, and DQB cDNA probes. Twenty-six patients had primarily chronic progressive MS and 74 had relapsing/remitting MS. The latter group included patients with a secondary progressive evolution of symptoms. Both clinical forms of MS were found to be associated with the DRw15,DQw6 haplotype. In addition, primarily chronic progressive MS was positively associated with the DQB1 restriction fragment pattern seen in DR4,DQw8, DR7,DQw9, and DRw8, DQw4 haplotypes, as well as negatively associated with the Taq I DQB1 allelic pattern corresponding to the serological specificity DQw7. Relapsing/remitting MS was positively associated with the DQB1 allelic pattern observed in the DRw17,DQw2 haplotype. These three DQB1 alleles are in strong negative linkage disequilibria with DRw15. The two susceptibility markers of each clinical form of MS act additively in determining the genetic susceptibility, as the relative risks for individuals carrying both markers roughly equal the sum of respective risks. Different alleles of the DQB1 locus defined by restriction fragment length polymorphisms contribute to susceptibility and resistance to primarily chronic progressive MS as well as to susceptibility to relapsing/remitting MS. The observed immunogenetic heterogeneity between the different clinical forms of MS favors the hypothesis that primarily chronic progressive MS and relapsing/remitting MS are two distinct disease entities.

Primarily chronic progressive and relapsing/remitting multiple sclerosis: two immunogenetically distinct disease entities.

Normal mouse spleen cells initiate synthesis of immunoglobulins in vitro in the presence of fetal calf serum and in the absence of specific antigen following the removal of regulatory suppressor(s) present in normal syngeneic mouse serum.

"Spontaneous" B cell activation due to loss of normal mouse serum suppressor.

Experiments were performed to study the mechanisms of the sensitivity of normal and neoplastic mouse cells to the cytotoxic action of humoral isoantibodies. A quantitative in vitro absorption procedure was used to compare the residua] hemagglutinin titer of a certain isoantiserum after the absorption of a known volume with known cell numbers. The cell numbers were adjusted so as to yield the same average total cell surface area. Lymph node cells and various solid and ascites tumor cells of strain A and C3H origin were used. It was found that different cell types differed with regard to their concentration of isoantigenic surface receptors. High correlation coefficients were found between the cytotoxic sensitivity and the concentration of isoantigenic receptors. It was shown that tumor cells which were partially or completely resistant to a single isoantiserum could be made sensitive to humoral isoantibodies provided that they were treated with mixtures of antisera, produced in different donor genotypes against the same target genotype, since each individual genotype reacts against a different antigen combination. This supports the concept that cytotoxic sensitivity is related to the surface concentration of reactive sites. Hematopoietic liver cells of newborn mice, which are completely resistant to isoantibodies, and spleen and lymph node cells of adult animals which are highly sensitive, were compared with regard to their ability to take up fluorescein-conjugated normal mouse serum in vitro. No differences were found between their visible pinocytotic uptake. Similar experiments were performed with various ascites tumor cells having different cytotoxic sensitivity. All tumor cells studied had a low pinocytosing ability which was not correlated to their sensitivity to humoral isoantibodies.

/pdf/quantitative-studies-of-the-sensitivity-of-normal-and-ye8ctldm5p.pdf

Quantitative studies of the sensitivity of normal and neoplastic mouse cells to the cytotoxic action of isoantibodies.

The ability of specifically immunized lymphoid cells to kill H-2 incompatible target tumor cells in tissue culture was shown to depend on the source of the lymphoid tissue (spleen versus lymph nodes). Marked cytotoxic effects were obtained with regional lymph node cells 7 to 10 days after primary immunization, whereas spleen cells from the same animals had little or no effect. Hyperimmunization did not decrease the cytotoxic efficiency of lymph node cells. Experiments were performed to test the possibility that the weak effect of spleen cells is a result of humoral antibody production, antagonizing the cell-bound immunity.

Humoral antibodies were cytotoxic in vitro in the presence of complement only. Their effect was manifested after 2 hours, whereas immune lymph node cells did not require complement and cytotoxicity was not expressed until 24 to 48 hours' incubation. Tumor cell cultures treated with specific humoral antibodies in the absence of complement became resistant to the cytotoxic effect of subsequently added immune lymph node cells, while no such protection was seen when normal serum was added. Thus, humoral antibodies led to an "efferent" inhibition of cell-bound immunity in vitro , in analogy with previous results in vivo .

/pdf/antagonistic-effects-of-humoral-isoantibodies-on-the-in-1xtffz272e.pdf

Erna Möller

Papers

Contact-induced cytotoxicity by lymphoid cells containing foreign isoantigens.

Primarily chronic progressive and relapsing/remitting multiple sclerosis: two immunogenetically distinct disease entities.

"Spontaneous" B cell activation due to loss of normal mouse serum suppressor.

Quantitative studies of the sensitivity of normal and neoplastic mouse cells to the cytotoxic action of isoantibodies.

Antagonistic effects of humoral isoantibodies on the in vitro cytotoxicity of immune lymphoid cells.