An attempt to eliminate platelet antibodies from antilymphocyte serum (ALS) by absorption with unwashed human erythrocytes.

After repeated, 10-20 absorptions of ALS with one-half of its volume of unwashed human erythrocytes, performed each time for 30 min. at room temperature, the titres of the undesired thrombo-agglutinins and thrombolysins can be reduced or these antibodies can be eliminated from ALS. After absorption anticomplementarity of ALS increases and the lymphocytotoxin titre declines. The optimal number of absorptions which would not reduce markedly the immunosuppressive activity of ALS remains to be determined.

[Cross-reactivity of HLA antigens of the donor and patient--a cause of unsuccessful thrombocyte transfusion in patients with HLA antibodies]. / Krízová reaktivita HLA-antigenu dárce a nemocného--prícina neúspechu transfúzí trombocytu nemocných s HLA-protilátkami.

In the microlymphocytotoxic test, specific HLA sera reacted positively with 74.74% of 99 examined samples of lymphocytes which failed to have corresponding HLA antigens yet contained HLA antigens cross-reacting with the given HLA antibody. In the serum of some patients HLA antibodies were established against HLA antigens cross-reacting with the antigens of the patient. The results show that it is not suitable to select thrombocyte and leukocyte donors on the basis of cross-reacting antigens. Therapy of patients with HLA antibodies may be clinically unsuccessful and even result in posttransfusion reaction. Patients who do not have HLA antibodies may develop them.

[Homozygosity or an undetermined antigen at the HLA locus--a problem for forensic experts]. / Homozygotie nebo neurcený antigen na HLA loku--problém soudních znalcu.

The author analyzed possibilities as regards the solution of paternity suits where paternity depends whether on the HLA 1 locus he is a homozygote or not. The authors presents a case of solution of a suit where the result depended on whether the man is a HLA-A2 homozygote or whether he has on the locus along with HLA-A2 also HLA-28 antigen.

[HLA-DR antigens and leukemias]. / HLA-DR antigeny a leukémie.

In 79 leukaemic subjects (42 with acute leukaemia, 37 with chronic myelosis) the authors assessed HLA-DR antigens 1-5.7 and their frequency. As compared with a group of 130 healthy subjects, no significant difference was found. The results do not support a possible relationship of HLA-DR antigens and leukaemia.

[Do monocytic agglutinins exist?]. / Existují monocytární aglutininy?

Twelve monocytotoxic sera were examined with 15 samples of monocytes, using the micromonocytotoxic and micromonoagglutination test. Weak agglutinins [+/-, +] were detected concurrently with cytotoxins only in 3 of 180 examinations at 37 degrees C. Monocytotoxins were found in 23.33% of reactions. From the results ensues that the existence of monocytic agglutinins is dubious.

[Isolation of monocytes from peripheral blood by gradient centrifugation on Lymphoprep in a hypertonic medium]. / Izolace monocytu z periferní krve centrifugací nad gradientem Lymphoprep v hypertonickém prostredí.

The author describes a new method of isolation of monocytes from peripheral blood, involving hypertonization of the blood and isolation of monocytes by centrifuging above a gradient of hypertonic Lymphoprep. The method makes it possible to obtain a monocyte suspension with at least 85% cells; there are no B lymphocytes in the suspension and the monocytes are not damaged. The quality of the thus obtained monocytes was tested by examination with monocytic sera and with HLA-DR antibodies, whereby the test was concurrently made with monocytes obtained by adherence and with B lymphocytes.

[HLA antigens on erythrocytes. II. A successful attempt to bind HLA serum antigens to erythrocytes]. / HLA antigeny na erytrocytech. II. Uspesný pokus o navázání HLA antigenu ze séra na erytrocyty.

In experiments in vitro it proved possible to bind HLA antigens in serum on erythrocytes after 4 to 48 hours incubation at 20 degrees C and 4 degrees C. With the period of incubation the amount of bound HLA antigens increased. The absorption and neutralization test revealed that the HLA antigen content of erythrocytes, thrombocytes and serum in a given subject is the same. HLA antigens present on the erythrocytes of some subjects were not removed from their surface even after nursing 15 times with saline. Based on the results of the experiment it is not possible to decide whether HLA antigens detected on red blood cells are an integral part of their membrane or whether they are bound to the red cells from plasma.

[HLA antigens on erythrocytes. III. Detection using the absorption and bromelin test]. / HLA antigeny na erytrocytech. III. Prukaz absorpcním a bromelinovým testem.

HLA antigens on erythrocytes were assessed concurrently by three tests--the indirect antiglobulin, two-stage bromelin and absorption test--using polyspecific and monospecific sera against HLA antigens A1, A2, B7, B8 and B12. Evidence obtained by the bromelin test was successful only in rare instances. The positivity of the absorption test increased with the number of saturations: after triple saturation of the HLA serum with erythrocytes of the same subject readings were obtained twice to three times more frequently than after single absorption. As compared with the indirect antiglobulin test, the positive erythrocyte reactions in the absorption test were less frequent: with polyspecific HLA sera in 36.97% as compared with 78.15%, with monospecific HLA sera in 63.64% as compared with 100%. The absorption test is a suitable method for detection of HLA antigens on erythrocytes, supplementing the indirect antiglobulin test.

[Recombination of HLA loci A, B and C found during examination in a paternity dispute]. / Rekombinace mezi HLA loky A a B, C zjistená pri resení sporu o otcovství.

The paper describes a case of the recombination of HLA-A and HLA-B, C loci appearing in one out of three children, found in a paternity dispute. The recombination referred to haplotypes inherited from the mother.

[HLA antigens on erythrocytes. An attempt to determine the origin of HLA-A, B antigens]. / HLA antigeny na erytrocytech. Pokus o zjistení puvodu HLA-A, B antigenu.

HLA sera were absorbed with erythrocytes with proved HLA antigens and blood cells with HLA-A, B antigens absorbed in vitro from serum. Both types of red cells were 5x to 15x washed in saline and exposed for two hours to the action of 15% chloroquine. The degree of absorption of HLA antibodies and the decline of the score were equal or slightly different after absorption with erythrocytes with own or quinked HLA antigens. Washing in saline or processing of blood cells with chloroquine did not affect the resulting absorption. From the results of the experiment ensues that the link of HLA-A, B antigens is equally firm blood cells isolated from the peripheral blood stream and blood cells where the antigens were linked to them in vitro from serum. This suggests that the view is justified that the majority of HLA-A, B antigens detected in blood cells is secondarily linked from plasma.

[HLA antigen typing in paternity disputes. Analysis of results]. / Typování HLA antigenu v paternitních sporech. Rozbor výsledku.

Typing of 38-43 HLA antigens ruled out paternity in 33.75% of 726 men suspected, during 198 revisions in 71.72%. In 63.57% the man was ruled out concurrently in loci HLA-A and B incl. in 79.33% where the man and child had all HLA-A, B antigens different. Locus HLA-A alone was ruled out in 8.01%, locus HLA-B in 28.17%; in a single case the man was ruled out in locus HLA-C only. By family investigations the man was ruled out in three of 11 cases. In two cases of paternity of twins the man was ruled out from paternity only of one child. In one case superfecundation was involved, confirmed in systems Duffy, Inv, Hp, AP, in the second case the man was ruled out from paternity of the second child in the Hp system. Male carriers of antigens of haplotypes frequent in the population were not ruled out from paternity in high percentages: with HLA antigens A1, B8 in 37.19%, A2, B7 in 19.54%, A2, B12 in 24.72%, A3, B7 in 37.14%.

[The effect of chloroquine on lymphocytic HLA-A and B antigens]. / K otázce úcinku chloroquinu na HLA-A, B antigeny na lymfocytech.

The author investigated the effect of 20%, 15%, 10%, 5%, 2.5%, 1.25% chloroquine on HLA-A,B antigens on lymphocytes and on the reaction HLA-A,B antigen-antibody. HLA-A,B antigens are not released by chloroquine from the lymphocyte surface and their expressivity does not change, as was revealed by examination of the micro-lymphocytotoxic and absorption test. Higher concentrations of chloroquine cause poly-reactivity of lymphocytes. A 20%, 15% and sometimes also 10% and 5% of chloroquine solution block the reaction HLA-A, B antigen--HLA antibodies. The reason for this is the destructive action of chloroquine on rabbit complement.

[An unusual case of paternity dispute resolved by examination of HLA antigens of a former wife, daughter and a sister of the deceased man]. / Neobvyklý prípad paternitního sporu, resený vysetrením HLA antigenu u bývalé manzelky a dcery zemrelého a jeho sester.

The author describes a peculiar case of a paternity suite where the accused died and his parents were also dead. Possible paternity was assessed by examination of HLA antigen of his ex-wife and daughter from his former marriage and in his sisters.

[The importance of recombination in resolving paternity disputes]. / Význam rekombinance pro resení paternitních sporu.

For evaluation of the results of HLA examinations in paternity disputes the objective finding that the man has HLA antigens which the child inherited from the father, is decisive. The controversy as regards heredity ensuing from assessment of HLA haplotypes of the man by family studies is caused by recombination. The author presents a case of this type.

[The importance of comprehensive immunohematologic tests in paternity disputes]. / Význam komplexního imunohematologického vysetrení pri paternitních sporech.

The authors describe four cases of paternity suits where the paternity was not ruled out according to HLA antigens and was ruled out in signs of erythrocyte, serum and enzyme systems, although the capacity of the HLA system was high, in two instances above 98%. From the results ensues that it is useful to supplement the examination by erythrocyte, serum and enzyme systems in those instances where HLA typing was performed first and the man was not ruled out.

[A rare case of paternity elimination based only on the HLA-C locus in the HLA system]. / Vzácný prípad vyloucení muze z otcovství v HLA systému jen na loku HLA-C.

The authors describe a rare paternity suit where paternity was ruled out in the HLA system only in locus HLA-C. The absence of HLA-Cw2 antigen on the lymphocytes was proved by the negative results confirmed by the same result on examination of the erythrocytes Ss system.

[An unusual paternity dispute case: classical methods did not rule out either of the two possible fathers]. / Neobvyklý prípad paternitního sporu: klasickými metodami nevyloucen zádný ze dvou mozných otcu.

The author described an uncommon case when neither of two men (former husband and another man) who were considered possible fathers of the child, could be ruled out as potential fathers based on examination of signs of 19 erythrocyte, serum and enzyme systems and 59 HLA-A, B, C antigens and their subtypes. The eliminating capacity of the examination was 99.86%; paternity was, based on probability calculation, practically proved (p = 99.9925% and 99.90%). According to statements of parties in the dispute the other man is the child's father. The author discusses perspectives of DNA examinations.

[Detection of HLA-A and HLA-B antigens on thrombocytes. Comparison of 3 methods]. / Prukaz HLA-A, B antigenu na trombocytech. Srovnání trí metod.

HLA antigens A1, 2, 9, 10, B5, 7, 8, 12, 35 were determined on 28 samples of healthy persons' thrombocytes by means of three tests; the microabsorption and immunofluorescence tests and the ELISA technique. Identification with the ELISA test was exceptional. Unlike the microabsorption test, the immunofluorescence test was employed to identify HLA antigens with only some sera as a rule. Identical positive results of both the microabsorption and the immunofluorescence tests were rather infrequent. The authors could not corroborate literary reports on difficulties in the identification of the antigens HLA-B8 and HLA-B12, certain problems were encountered in the case of the HLA-B7 antigen only. A discussion is presented of the causes of the different results of each of the tests, and of the relevance of the results for transfusions of platelet concentrates chosen on the basis of lymphocytic cross-matching.

[Is the fluorescence cytotoxicity test for the determination of HLA-DR antigens more sensitive than the classical microcytotoxicity test?]. / Je fluorescencní cytotoxický test v urcování HLA-DR antigenu citlivejsí nez klasicky mikrocytotoxický test?

The authors compare the sensitivity of the testing methods for determining HLA-DR antigens on B-lymphocytes and monocytes (classical eosin technique) with the fluorescence cytotoxic tests (FCT). The assessment of results of DR specificity determination in blood donors and patients with blood disease showed comparability of the tests. The rate of concordance ranged from 88-89%. The paper discusses the advantages and disadvantages of each method.

[An unusual case of eliminating a man as the father of twins]. / Neobvyklý prípad vyloucení muze z otcovství k dvojcatum.

The authors describe an uncommon case when despite the great eliminating capacity of the HLA system a man in this system was eliminated only from paternity of one twin; as to the other twin, surprisingly he was eliminated in the Hp serum system.