Ten-year follow-up of HIV infection in a haemophiliac cohort.
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Acquired Immunodeficiency Syndrome↗
Biomedical subjects
G Janossy
Publications and source records attributed to G Janossy.
The development of primed/memory CD8+ lymphocytes in vitro and in rejecting kidneys after transplantation.
In normal individuals, 80 +/- 5% of circulating CD8+ T cells express CD45RA, and 20 +/- 7% of these cells express CD45R0 antigens. After activation, CD8+ cells expressing CD45RA decrease to 56-67% while those expressing CD45R0 increase to 38-67%. Although precursors of alloantigen-specific cytotoxic T cells were found in both CD8+, CD45RA+ and CD8+, CD45R0+ subsets, the specific effector cells were exclusively CD8+, CD45R0. Allospecific cytotoxic CD8+ clones were also entirely CD45R0+. A lectin-dependent cytotoxic (LDC) assay unmasked a hierarchy of killing after alloactivation which was CD8+, CD45R0+ greater than CD8+, CD45RA+ greater than CD4+, CD45R0+ greater than CD4+, CD45RA+. The phenotype of CD8+ T cells in rejecting kidneys was similar to in vitro alloantigen-activated CD8+, CD45R0+ cells and cytotoxic CD8+ clones. Firstly there was an increase in the relative proportions of CD45R0+ (60 +/- 8%) and a decrease in CD45RA+ (35 +/- 10%) CD8+ cells relative to circulating CD8+ subsets. In the rejecting grafts, 34 +/- 9% of the CD8+ cells were also DR+ indicative of recent activation. Furthermore, 16% (range 4-35%) of rejecting CD8+ cells were Ki67+ suggesting that these cells were proliferating. Finally, 17% (range 4-53%) of T cells in rejecting kidneys simultaneously expressed both CD45RA and CD45R0 markers. These results show that in vitro alloantigen-activated CD8+, CD45R0+ cells represent a primed/memory cytotoxic population. In addition, they provide indirect evidence that a proportion of CD8+ cells in rejecting kidneys were actively switching from a naive to a memory phenotype in vivo in a manner analogous to that in vitro.
Antibodies, Monoclonal↗
Mononuclear cells in glomeruli and cytokines in urine reflect the severity of experimental proliferative immune complex glomerulonephritis.
Immunohistochemical methods were used to investigate the role of macrophages in the progression of proliferative immune complex glomerulonephritis. The mononuclear cell component of glomerular inflammation was analysed in three different stages of chronic serum sickness, each of which was clearly distinguished by criteria of kidney function. Urinary excretion of the macrophage secretory products interleukin-1 and tumour necrosis factor was also evaluated in relation to the functional severity of kidney disease. T lymphocytes and macrophages began to accumulate in glomeruli at the onset of proteinuria, but not before. Urinary excretion of interleukin-1 also began with proteinuria. Proteinuria increased in direct correlation with increases in the number of glomerular macrophages. Development of the most severe stage of glomerulonephritis, characterized by cachexia, declining kidney function, and necrotizing glomerular pathology, was accompanied by the disappearance of T cells from glomeruli and the expression of highly abnormal phenotypes by most macrophages. In addition, there was a switch from urinary excretion of interleukin-1 to excretion of tumour necrosis factor. The progression of proliferative immune complex glomerulonephritis was associated with qualitative as well as quantitative changes in glomerular macrophage populations. Differentiation and/or activation of those glomerular macrophages may have resulted from local T cell-mediated immunoregulation. Measurements of urinary cytokine excretion provided a reliable means of monitoring disease progression. The local action of tumour necrosis factor probably contributed to declining kidney function in the most severe stage of disease.
Animals↗
The detection of residual acute lymphoblastic leukemia cells with immunologic methods and polymerase chain reaction: a comparative study.
In this study we applied double color immunofluorescence analysis and polymerase chain reaction (PCR) amplification of rearranged TCR delta genes for detecting residual leukemia in the posttreatment bone marrow (BM) samples taken from four patients in morphological remission. In three of these patients (nos. 1-3; T-ALL) a combination of CD3 and anti-TdT antibodies (Abs) was used to identify residual blasts while in patient 4 (B lineage ALL) the combination CD13/TdT served to detect residual disease. Two rounds of PCR primed by nested amplimers were carried out to prepare clonospecific probes from presentation DNA and to investigate the follow-up samples. In patients 1 and 2 no cCD3+/TdT+ cells were seen posttreatment, but PCR amplification of the TCR V delta 1-D-J delta 1 region revealed residual disease in both patients. Patient 1 underwent allogeneic BM transplant (BMT) 8 months after diagnosis and is well 3 months post-BMT while patient 2 relapsed 12 months after presentation. In patient 3 the remission samples investigated 2 and 3 months after diagnosis did not contain cCD3+/TdT+ cells, but in the sample collected at 4 months a few such cells (0.0001-0.001%) could be detected. In the same sample, PCR amplification of the TCR V delta 2-D-J delta 1 region indicated the presence of 10(-4)-10(-3) residual leukemic cells. These findings predicted full morphological relapse which occurred 2 months later. In patient 4 CD13/TdT double positive cells were clearly seen 2 and 3 months after presentation. PCR amplification of the V delta 2-D delta 3 recombination also revealed residual blasts when applied to one of such "remission" samples. After further remission induction treatment, no immunologic evidence of residual disease was detected. This patient received an allogeneic BMT 8 months after diagnosis and is disease free 4 months after BMT. Our data indicate that both double color immunofluorescence and PCR analysis offer powerful tools to study residual leukemia and highlight the advantages as well as the potential limitations of each technique.
Antigens, CD↗
Lymphatic tissue changes in AIDS and other retrovirus infections: tools and insights.
Based on immunohistochemistry, in situ hybridization, and electron microscopy, lymphatic tissue changes in human immunodeficiency virus (HIV) and other retroviral infections represent different stages of a dynamic process progressing from hyperplasia to atrophy. The germinal centers (GC) function early as a virus reservoir in both HIV and feline leukemia virus infection, which also produces lymphadenopathy, severe immune impairment, and death from opportunistic infections. Core proteins of HIV can be detected on the surface of follicular dendritic cells, electron microscopy reveals cell-free HIV and virus replication in the same location, and in situ hybridization shows that the majority of cells with mRNA of HIV can be found in germinal centers (GC). Double immunohistochemical labeling of lymphocyte populations suggests that one of the most important events in HIV lymphadenitis with explosive follicular hyperplasia is the accumulation of CD8+CD45R0+ lymphocytes in the lymph nodes. Their clustering in the vicinity of the FDC network could play a key role in disintegration of GC and lymphocyte depletion as the disease progresses.
Acquired Immunodeficiency Syndrome↗
Phenotypic, genotypic, cytochemical, and ultrastructural characterization of acute undifferentiated leukemia.
During the diagnostic investigation of 750 acute leukemias, nine cases were morphologically, cytochemically, and phenotypically undifferentiated. In seven of these cases the blasts were class II+, CD34+ and TdT+, in one were class II+, TdT+, CD7+ while in the remaining leukemia blasts expressed class II only. Cytoplasmic and membrane CD22, CD3, CD13, and Ig as well as membrane CD19, CD10, CD37, CD2, CD33, CD14, glycophorin C, and CD61 were absent. The further characterization of these rare leukemias yielded the following results. The TCR-beta, -gamma and -delta genes were in germline configuration in seven cases studied while IgH genes were rearranged on both alleles in two cases and germline in the other five. By ultrastructural analysis peroxidase activity was detected on unfixed cells in a minority of blasts from four of seven cases. In two of the peroxidase-positive cases a small proportion of blasts also reacted with an anti-myeloperoxidase monoclonal antibody. In one of the peroxidase-negative cases, 7% of blasts were labeled by the antibody, suggesting the presence of peroxidase in its proenzyme form. Importantly, the two cases with Ig gene rearrangements did not have cytochemically or immunologically detectable peroxidase. Three of the nine patients were treated as ALL while six received AML chemotherapy. In five patients complete remission was achieved while the other four died from infections during remission induction. Four patients are still in remission 7, 12, 24, and 30 months after diagnosis while one patient relapsed after 12 months. In conclusion, we have characterized the genotypic and ultrastructural features of subtype of acute leukemia in which blasts expressed immaturity markers and lacked lineage associated antigens. In contrast to previously reported "unclassifiable" cases, the leukemias were phenotypically homogeneous and showed a good response to chemotherapy.
Acute Disease↗
Characterization of a human T cell-specific chimeric antibody (CD7) with human constant and mouse variable regions.
A chimeric human-mouse anti-T lymphocyte mAb (CHT2; SDZ 214-380) has been constructed by cloning the variable region exons of both the L and H chains from the murine hybridoma RFT2 which have CD7 specificity and reactivity with a 40-kDa Ag. The variable L chain exon was joined to the human C kappa, and the variable H chain exon was joined to the human IgG1 region exon encoding the human allotype nGlm(z), nGlm(a). The gene constructs were introduced by electroporation into SP2/0, a non-Ig-producing murine myeloma. The identical tissue reactivity of the newly made CHT2 and the original murine RFT2 mAb (CD7) was confirmed by blocking experiments as well as by immunohistology and flow cytometry. Because this new mAb may have clinical use, the CD7 Ag expression of T lineage cells has also been quantitated in double and triple immunofluorescence assays in combinations with mAb to restricted forms of leukocyte common Ag that designate unprimed (CD45R+) and primed T lymphocyte populations (UCHL1+). CHT2 shows very strong reactivity with large thymic blast cells and cortical thymocytes from which T-ALL originates. Strong staining is seen on CD45R+ unprimed "virgin" T lymphocytes, whereas the expression on UCHL1+ primed "memory" cell types is weaker unless these cells are reactivated by mitogens or Ag. Thus CHT2 may spare a substantial population of resting memory T cells which is relevant to its potential therapeutic use. In addition the chimeric antibody had a greater in vitro antibody dependent cytotoxicity and a prolonged half-life (4.2 to 5.0 days) in Rhesus monkeys.
Amino Acid Sequence↗
The expression of T cell receptor-associated proteins during T cell ontogeny in man.
The expression of TCR-associated molecules was examined in human fetal and postnatal tissues. From gestational wk 7 onward in the fetal liver, putative prothymocytes have been identified with cytoplasmic CD3 positivity (cCD3+). These immature cells are TdT- and do not express membrane CD3 (mCD3-) or TCR beta identified by beta F1, but show CD7 and CD45 positivity without CD1, CD2, CD5, CD4, CD8, CD10, and class II Ag. Their high proliferative activity is indicated by greater than 85% Ki67 positivity. After the 10th wk, beta F1+, mCD3+ cells also appear in the liver and these are mostly Ki67- but no TCR gamma delta-bearing cells can be identified at such an early stage of extrathymic development. In the mCD3- TdT-fetal thymus (10 1/2 to 18th wk) cCD3+, mCD3- CD1-blasts proliferate (Ki67+) and lack TCR-beta or TCR-gamma delta. The TdT-, CD1+ cortical thymocytes develop into TCR-beta + and WT31-positive (TCR-alpha beta +) cells. Subsequently TdT-positive thymocytes become detectable around 19 to 20 wk, and in such glands the peak of proliferative activity is seen among TdT+, cCD3+ cells which appear to acquire, in a regular sequence, cytoplasmic beta F1 (TCR-beta), mCD3, and TCR-alpha beta (WT31 positivity) together with the loss of TdT and Ki67 positivity. A newly described transitional population of cells is TdT-, beta F1+ but exhibits no detectable WT31 positivity. These cells correspond to the CD1+, mCD3+ thymocytes and are probably the targets of thymic selection. The cells of the TCR-gamma delta lineage, detected by mAb TCR-delta-1 and delta TCS1, are rare (0.02 to 0.5%) among thymocytes from gestational wk 10 1/2 onward through the whole span of thymic development, but these cells include a proportion (18 to 59%) of cells expressing CD1 Ag, suggesting that these TCR-gamma delta cells differentiate in the thymus. Among the CD1+, TCR-gamma delta + thymocytes, no TdT positivity can be detected.
Adolescent↗
Kinetics of T lymphocyte development.
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Animals↗
The expression of T-cell receptor-associated proteins in normal and leukaemic immature T cells.
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Cell Differentiation↗
The epitope specificity and tissue reactivity of four murine monoclonal anti-CD22 antibodies.
The CD22 antigen is expressed on the surface of normal human B cells and some neoplastic B cell lines and tumors. Previous cross-blocking studies using a panel of monoclonal anti-CD22 antibodies have defined four epitope groups, termed A-D. In the present studies, we have further dissected the epitopes recognized by four monoclonal anti-CD22 antibodies using immunoprecipitation and cross-blocking techniques, immunofluorescence analyses with a variety of cell lines, and immunoperoxidase analyses of 36 normal human tissues. Two of the antibodies, HD6 and RFB4, have been described previously, and two, UV22-1 and UV22-2, are described in this report. Our studies indicate that the four monoclonal antibodies show unexpected complexities in their reactivity with CD22+ and CD22- cells and their reactivity with solubilized CD22 molecules. The four antibodies, which recognize epitopes defined previously as CD22-A and CD22-B, further subdivide these epitope clusters into four determinants, A1, A2, B1, and B2. Furthermore, only two of the antibodies, RFB4 and UV22-2, are B cell-specific. In summary, our data indicate that RFB4 and UV22-2 would be the antibodies of choice for constructing immunotoxins to treat B cell tumors.
Animals↗
Prediction of progression to AIDS by analysis of CD4 lymphocyte counts in a haemophilic cohort.
Serial CD4 lymphocyte counts were recorded in 112 anti-HIV-positive haemophiliacs who were followed for up to 8 years after seroconversion. The patients remained at low risk of developing AIDS until their CD4 lymphocyte count fell to 0.25 X 10(9)/l. From this point, the risk increased as their count approached zero. Using this result and on the assumption (which is evaluated) that the underlying trend over time in CD4 lymphocyte counts is linear, the predicted rate of progression to AIDS was calculated for the cohort. It was estimated that 73% (95% confidence limits 60-86%) of the cohort will develop AIDS within 15 years of HIV-seropositivity. During 8 years of follow-up, this cohort had shown similar rates of progression to AIDS to other cohorts--haemophilic and otherwise--suggesting that this estimate may well have general applicability. The method described could be used to plan the provision of health-care resources for groups of anti-HIV-positive patients as it allows the number of new cases of AIDS to be predicted year by year, even when the patients' dates of seroconversion are unknown.
Acquired Immunodeficiency Syndrome↗
The relevance of induced class II HLA antigens and macrophage infiltration in early renal allograft biopsies.
Using a panel of monoclonal antibodies, immunohistological analysis was performed on frozen sections taken from 14 peritransplant renal biopsies and 42 biopsies taken 6 +/- 2 days posttransplantation. The following parameters were examined: tubular expression of HLA-DR, DP, and DQ and infiltration with T lymphocytes and macrophages. Of the 42 posttransplant biopsies, 26 were diagnosed as rejecting and 16 as nonrejecting according to clinical and histopathological criteria. HLA-DR antigens were strongly expressed on 8 of 14 peritransplant biopsies, 23 of 26 rejecting biopsies and 13 of 16 nonrejecting biopsies. Tubular expression of HLA-DP and DQ was weak or absent. In the rejecting biopsies there was a significantly increased infiltrate of T lymphocytes of all phenotypes and of macrophages when compared with the nonrejecting biopsies. Graft outcome was invariably favorable in the nonrejecting group, with no graft losses in the first posttransplant year. There were 4 graft losses in the rejecting group, all due to rejection, and further analysis revealed that all 4 had macrophage-dominated infiltrates in their early allograft biopsies. We conclude that immunohistological analysis of early allograft biopsies provides an accurate prognosis of subsequent graft acceptance or rejection and that early macrophage infiltration is a poor prognostic sign.
Antibodies, Monoclonal↗
The reliability of cytoplasmic CD3 and CD22 antigen expression in the immunodiagnosis of acute leukemia: a study of 500 cases.
Current views about the origin of acute lymphoid leukemia (ALL) emphasize the importance of maturation arrest at a precursor cell level. Recently, the CD22 antigen has been identified in the cytoplasm of normal bone marrow-borne immature B lineage cells, while the CD3 antigen (epsilon chain) has been detected within normal immature thymic blasts. In the first part our study performed on 100 cases of known acute leukemias, the expression of such cytoplasmic molecules, referred to as cCD22 and cCD3, was analyzed together with their appearance in the leukemic cells' membrane (mCD22 and mCD3). The presence of cCD22 in B-lineage ALL and that of cCD3 in T-ALL has indeed fully confirmed the diagnosis reached by other markers, and mCD22 and mCD3 were expressed on only a few cases of B- and T-lineage ALL, also revealing a degree of developmental asynchrony within leukemic blasts. In the subsequent analysis both cCD22 and cCD3 have been included in a standard panel of diagnostic reagents applied on 500 consecutive cases of acute leukemia. Here the aim was to analyze both the diagnostic precision of individual markers and the heterogeneity of various leukemic types in terms of the expression of membrane and intracellular antigens and their cytochemical features (Sudan Black B and esterases). It has been found that cCD22 and cCD3 are exquisitely specific for B-precursor ALL (TdT+, CD19+) and T-ALL (TdT+, CD7+), respectively, while both markers are absent in acute myeloblastic leukemia (AML) and acute myelomonocytic and monocytic leukemia (AMML/AMoL). These observations contrast the findings which demonstrate that 31% of cases among nonlymphoid acute leukemia (including AML and AMML) express CD7 and/or TdT. The study of myeloid antigens detected by CD13, CD33, and CD14 is also informative and complementary, both in diagnosing and subdividing the AML and AMML/AMoL groups. The peculiar main observation of this study is that only with the combined use of these markers in a microplate assay for membrane antigens, followed by double staining for intracellular antigens such as terminal deoxynucleotidyl transferase, cCD3, cCD22, c mu heavy chain, and T cell receptor beta, it is possible to safely establish the lineage affiliation and subgrouping of virtually all acute leukemias. Among these cases are those with aberrant combinations of markers, including 14% of B-lineage ALL (cCD22+,CD19+,TdT+) and a single case T-ALL (cCD3+,CD7+,TdT+), which exhibit CD13 and/or CD33 antigens, cases with mixtures of ALL and AML blasts, and 1.2% of acute leukemias which lack lineage affiliation and can be regarded as acute undifferentiated leukemia.
Antigens, CD↗
Cellular events during memory T-cell activation in vitro: the UCHL1 (180,000 MW) determinant is newly synthesized after mitosis.
After T-cell activation, a switch of leucocyte common antigen (LCA) expression occurs which is detected by the disappearance of CD45R and the appearance of UCHL1 positivity. Upon activation of CD45R+ T cells by phytohaemagglutinin (PHA), the IL-2 receptor (IL-2R) develops at 15 hr and blast cells enter the S phase at 20 hr, while remaining UCHL1-. At 40 hr, mitotic cells continue to express CD45R determinants, but weak UCHL1 reactivity is now detectable on 15-20% of these cells. By inhibiting the separation of daughter cells with cytochalasin B, it can be demonstrated that the UCHL1 antigen is newly synthesized in the Golgi apparatus before insertion into the membrane after the first mitosis. At 65 hr after activation, 80% of proliferating cells are UCHL1+. T cells undergo a similar development during allogeneic activation, and specific alloresponsive cells, tested in secondary mixed lymphocyte reaction (MLR), become greatly enriched among the proliferating UCHL1+ cells. Conversely, after a primary MLR, residual CD45R+ T cells are unresponsive to the original stimulus but can still proliferate in response to unrelated alloantigens. These results clarify both the time-course of the acquisition of UCHL1 antigen during the proliferative cycle of activated T cells, and indicate the shift of antigen responsive T-cell populations from the CD45R+ to the UCHL1+ pool.
Cells, Cultured↗
T10B9.1A-31 anti-T-cell monoclonal antibody: preclinical studies and clinical treatment of solid organ allograft rejection.
T10B9.1A-31 is an immunoglobulin Mk (lgMk), CD3b class, murine monoclonal antibody. It is not itself mitogenic, but it partially blocks mitogenesis produced by concanavilin-A, phytohemagglutinin, and a soluble antigen cocktail and is lytic for human peripheral blood T lymphocytes (PB-T) in the presence of fresh autologous human complement and rabbit complement. It reacts with a monomorphic epitope found on all mature PB-Ts that modulates in vitro and in vivo with the CD3/T-cell antigen receptor (TCR) complex, but unlike OKT3, which reacts with CD3 proteins, T10B9.1A-31 reacts with an epitope of the TCR alpha/beta heterodimer. Immunoprecipitation of HPB-ALL with T10B9.1A-31 or OKT3 revealed that T10B9.1A-31 does not react with the 20-to 26-kd polypeptides that compose CD3, and T10B9.1A-31 failed to bind to the PEER cell, which is CD3 gamma/delta positive, TCR alpha/beta negative. Phase 1 clinical trials demonstrated that T10B9.1A-31 caused a rapid decrease in PB-Ts and no toxic effects other than fever, chills, rigors, and transient hypotension. These side effects were absent in methylprednisolone (MP)-pretreated patients. Phase II studies revealed that T10B9.1A-31 reversed steroid refractory acute rejection in three of five renal allografts treated with cyclosporine (CyA) and prednisone (P) maintenance immunosuppression, reversed steroid and polyclonal antisera refractory rejection in three cardiac allograft recipients receiving CyA/P, and attenuated acute rejection in three of three renal patients receiving baseline azathioprine/prednisone (AZA/P) when used as primary therapy. Rerejection was not seen when patients received CyA baseline immunosuppression but was seen in patients maintained on AZA/P OKT3 was efficacious in treating rejection following T10B9.1A-31 therapy. Side effects of T10B9.1A-31 appears to reverse acute rejection crisis effectively in renal and cardiac transplantation with a low incidence of rerejection in CyA-maintained patients and with fewer, milder side effects. Sequential therapy with OKT3 is possible because OKT3 and T10B9.1A-31 are of different isotypes and idiotypes.
Animals↗
Lack of specialized antigen-presenting cells (IDCs) in donor heart, kidney, liver, and lung at transplantation.
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Antigen-Presenting Cells↗
The tissue distribution of T lymphocytes expressing different CD45 polypeptides.
The distribution of T lymphocytes expressing the different polypeptides of the leucocyte common antigen (LCA) family detected by CD45R and UCHL1 antibodies has been studied in normal lymphoid tissues. In the thymus most cortical thymocytes express UCHL1 and co-express CD4 and CD8. The more mature membrane CD3+ (mainly medullary) T cells are heterogenous and may express both UCHL1 and CD45R weakly or be restricted to display CD45R or UCHL1 alone. In the medulla both the CD45R+ and UCHL1+ subpopulations contain single positive CD4 and CD8 cells. In tonsils, germinal centre T cells are almost exclusively UCHL1+, CD4+ and a proportion also express HNK-1 (Leu 7) antigen. In the paracortical areas approximately equal numbers of CD45R+ and UCHL1+ cells are found but these separately occupy nests of cells containing one or the other type. Again, both CD45R+ and UCHL1+ cells include single CD4+ and CD8+ lymphocytes. A small proportion (less than 5%) of strongly CD45R+, UCHL1+ double-stained cells are also seen, and these probably represent recently activated lymphocytes. In the gut, small clusters of such strongly double-labelled cells are in the submuscular mucosae while cells of the lamina propria are almost exclusively UCHL1+. Many intra-epithelial lymphocytes are only weakly positive or negative for UCHL1 and appear to be CD45R-. These results are consistent with the view that expression of different CD45 polypeptides identifies successive stages of thymocyte-T-cell maturation and that following their thymic education, unprimed T lymphocytes are CD45R+, while primed memory T cells are UCHL1+. These populations occupy different microenvironments.
Antigens, Differentiation↗