T-cell clonotype assay as immunological monitoring in renal and bone marrow transplantations.
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Bone Marrow Transplantation↗
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Advances in the immunological monitoring of childhood acute lymphoblastic leukaemia.
In children with acute lymphoblastic leukaemia (ALL), measurements of minimal residual disease (MRD) during therapy provide crucial information about the response to treatment and the risk of relapse. Flow cytometry is a practical and widely applicable tool for monitoring MRD in patients with ALL. This approach is based on the identification of immunophenotypes expressed by leukaemic cells but not by normal lympho-haematopoietic cells in bone marrow and peripheral blood. These phenotypes can identify one leukaemic cell among 10 000 normal cells and are currently applicable to at least 90% of patients with ALL. A strong correlation between flow cytometric measurements of MRD during clinical remission and treatment outcome has been demonstrated, suggesting that these assays should be incorporated into treatment protocols.
Child↗
Use of monoclonal antibodies to T-cell subsets for immunologic monitoring and treatment in recipients of renal allografts.
Using monoclonal antibodies and flow cytometry, wer serially monitored lymphocyte subpopulations in renal-allograft recipients treated with either conventional immunosuppression or a monoclonal antibody. In 29 patients given conventional suppression, highly significant correlations between changes in T-cell subsets and rejection were noted. Normal or elevated ratios of OKT4 (helper/inducer) to OKT8 (suppressor/cytotoxic) cells were associated with rejection unless the donor was HLA identical or the total number of T cells was extremely low. In patients with low ratios, rejection seldom occurred. Two patients treated with OKT3 monoclonal antibody for acute rejection had rapid disappearance of OKT3-reactive cells from the peripheral blood and prompt reversal of rejection. The use of monoclonal antibodies allows the precise determination of changes in T-cell subsets and promises the development of therapeutic protocols that can be designed to manipulate selected lymphocyte populations.
Antibodies↗
Immunologic monitoring in lung allograft recipients.
To identify patients with increased risk of chronic lung allograft rejection, we assessed the utility of an in vitro biopsy-derived lymphocyte growth assay and serum anti-HLA antibody screening as a complement to currently available methods of monitoring lung allograft recipients. Lymphocyte growth assay was performed on bronchoscopic fragments of tissue cultured in medium with rIL-2. Seventy-nine biopsies from 31 lung transplant recipients were tested by lymphocyte growth assay, and results were correlated with histopathology findings. Positive lymphocyte growth was found in 12/26 (46%) episodes of acute rejection, 5/44 biopsies without rejection (11%), and 0/9 episodes of bronchitis. Positive lymphocyte growth was seen in 7/16 (44%) grade A1 rejections and in 5/10 (50%) grade A2 rejections, as opposed to only 5/44 (11%) grade A0 (no rejection) biopsies (P < 0.01 for both A1 and A2 with respect to A0). Actuarial probability of remaining free from obliterative bronchiolitis (OB)* tended to be higher in patients who did not exhibit lymphocyte growth in biopsies. Sequential samples of sera obtained at the time of the biopsy were screened for lymphocytotoxic anti-HLA antibodies. Twenty-two of 44 recipients (50%) developed anti-HLA antibodies during the first postoperative year, exhibiting greater than 10% reactivity to an HLA reference panel of lymphocytes in four or more consecutive serum samples. Actuarial survival of lung allograft recipients with anti-HLA antibodies (n = 22) was lower than in those without anti-HLA antibodies (n = 22; P = 0.03). Of the 22 antibody producers, 7/12 died as a consequence of OB. Of the 22 non-antibody-producers, 1/2 deaths occurred as a consequence of OB. Anti-HLA antibodies were present in 9/11 instances of OB (82% sensitivity) and in 13/33 patients without OB (61% specificity; P = 0.03). These data indicate that lung transplant recipients with positive lymphocyte growth and anti-HLA antibodies are at an increased risk of chronic allograft rejection.
Antibodies↗
Clinical value of immunological monitoring of minimal residual disease in acute lymphoblastic leukaemia after allogeneic transplantation.
In this study, we used multiparameter flow cytometry to quantify minimal residual disease (MRD) in 165 serial bone marrow samples from 40 patients diagnosed with acute lymphoblastic leukaemia (ALL) who underwent allogeneic stem cell transplantation (allo-SCT) from siblings (n = 34) or unrelated donors (n = 6). Samples were prospectively taken from 24 patients before starting the conditioning regimen, at days +30, +60 and +90 and subsequently every 2-3 months. Samples from 16 patients in complete remission (CR) after allo-SCT were taken at least twice. Six of 24 patients harboured MRD (0.2-10% of mononuclear cells) at transplant and 18 were negative. Estimated disease-free survival for the MRD+ and MRD- groups at transplant was 33.3% and 73.5% respectively (P = 0.03). During follow-up, increasing MRD levels were detected in nine patients, a finding that preceded marrow relapse by 1-6 months. Two patients with stable low MRD levels remained in CR. When we used flow cytometry to test the effect of donor leucocyte infusions (DLI) in six patients, we observed that the only sustained remission was achieved when DLI was applied prior to overt relapse. We conclude that MRD by flow cytometry can rapidly assess tumoral burden before transplant to predict outcome, and can be clinically useful for the timing of DLI for increasing levels of leukaemia after transplant.
Adolescent↗
Immunological monitoring of a patient with Guillain-Barré syndrome successfully treated with plasma exchange.
A patient with Guillain-Barré syndrome was successfully treated by plasma exchange (without any other therapy), and the relationship between changes in his peripheral blood lymphocyte subpopulations and the clinical improvement induced by plasma-pheresis was studied. At the recovery, the most interesting findings were an increase in T suppressor lymphocytes, and in Fc receptor-bearing lymphocytes (above normal values), together with a decrease in B lymphocytes. In this patient, plasma exchange not only removed toxic humoral factors, but also restored a normal balance among peripheral lymphocytes.
Aged↗
Immunological monitoring of bone marrow transplant recipients. I. Major leucocyte subclasses in the blood.
In the recipients of an allogeneic HLA-identical sibling transplant the blood leucocytes were reconstituted within 3 to 4 weeks but the level of lymphocytes remained low throughout the observation period of 20 weeks. Of the different lymphoid cell subsets, the large granular lymphocytes (LGL) reconstituted fastest, followed by DR-expressing lymphocytes. The reconstitution was accompanied by a significant lymphoid blastogenesis in the blood. The frequency of OKT4 and OKT8 lymphocytes was initially low; the number of OKT8-positive lymphocytes reached normal levels by the 6-8th week whereas the number of OKT4-positive lymphocytes and, consequently, the OKT4/8 ratio remained low. The responses to the T mitogens, phytohaemmagglutinin and concanavalin A, were strongly suppressed. Only a few significant changes were observed before and during acute graft-versus-host disease (aGVHD): the frequency of LGL, lymphoid blasts and OKT8-expressing lymphocytes was depressed before aGVHD and the frequency of lymphoid blasts remained depressed throughout the episode. We conclude that reproducible changes occur in the leucocyte subset frequencies during reconstitution, but the characteristic changes prior to and during aGVHD are not particularly prominent and hardly of diagnostic use.
Adult↗
Is the determination of the soluble interleukin-2 receptor after application of interleukin-2 receptor antibodies still appropriate for immunological monitoring after renal transplantation?
INTRODUCTION: The use of monoclonal antibodies against the alpha-chain of the membrane-bound interleukin-2 receptor (IL-2Ralpha) as immune suppressants causes characteristic changes in the levels of soluble interleukin-2 receptor (sIL-2R) in serum and urine. METHODS: 38 kidney transplant patients were included in this study. 28 of them received an induction therapy with the IL-2R antibody basiliximax (Simulect) in addition to standard immunosuppression, 10 patients constituted the control group. RESULTS: Time courses of sIL-2R levels of Simulect patients with and without complications after transplantation have been compared. It turned out that of a total of 18 cases with complications 15 cases could be identified by their elevated sIL-2R levels, which corresponds to a sensitivity of 83%. CONCLUSION: Acute rejection, CMV infection, extrarenal bacterial infection and pyelonephritis in the transplant all cause a significant increase of the sIL-2R level even after application of Simulect.
Adult↗
Immunologic monitoring during combined radiochemoimmunotherapy in inoperable lung cancer.
49 patients with advanced lung cancer underwent a combined treatment including radiochemoimmunotherapy. Before and at the end of the combined treatment, the immune status of the patients has been evaluated by testing the immunoreactivity to PPD in vivo and the phytohemagglutinin lymphocyte response and the rosette-forming cells in vitro. While the in vitro tests were not significantly modified by immunostimulation, as regards the immunoreactivity to PPD, we observed a positive conversion at the end of the treatment in 11 out of 25 patients who showed a negative reaction before therapy. All the patients who converted from negative to positive showed also a good response to therapy with regression or no progression of the tumor. Survival curves confirmed the prognostic value of PPD reactivity.
BCG Vaccine↗
Immunological monitoring in systemic lupus erythematosus.
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Adolescent↗
Immunologic monitoring of the cardiac transplant patient.
Peripheral blood lymphocyte morphology and cell surface markers were repeatedly evaluated in 49 orthotopic cardiac transplant patients over a period of three years to determine the utility of these parameters in predicting an episode of acute cardiac rejection. Lymphocytes were measured with a calibrated microscope and termed "activated" if greater than 10 mu in diameter. If a patient demonstrated greater than 30 activated lymphocytes/cu mm, the same lymphocytes were stained with monoclonal antibodies directed to T-cell subsets and B cells. These findings were retrospectively correlated with endomyocardial biopsy results. Absolute numbers of T-cell subsets and B cells were analyzed via Student's t test to identify significant differences during acute cardiac rejection. Of 347 biopsy specimens, 47 demonstrated histologic features of acute cardiac rejection. Simultaneous immune activation of lymphocytes occurred with 33 of 47 samples, while another 51 episodes of lymphocyte activation were detected without acute rejection. No statistically significant differences in absolute numbers of T-lymphocyte subsets were noted at the time of acute rejection.
Acute Disease↗
Diagnosis of rejection in the allografted rat lung: using monoclonal antibodies to T cell subsets for immunologic monitoring.
Early diagnosis of rejection and timely immunosuppression are absolutely important in clinical lung transplantation. We studied surface markers of peripheral blood lymphocytes (PBL), graft infiltrating lymphocytes (GIF) and bronchoalveolar lavage fluid (BALF) in a rat using flow cytometric monitoring to diagnose rejection. Left lung transplantation was performed on Brown Norway (BN) rats and Lewis (LEW) rats in the following groups; Group 1: LEW-LEW (isograft), Group 2: BN-LEW (allograft; no immunosuppression), Group 3: BN-LEW (allograft; treated with Cyclosporine A at a dose of 15 mg/kg/day i.m.). In each group, rats were killed 3, 5, 7 days postoperatively (n = 6 on each day). Monoclonal antibodies investigated in this study were W3/25 (anti-helper T lymphocyte), OX8 (anti-suppressor/cytotoxic T lymphocyte), and OX39 (anti-interleukin 2 receptor). Histological classification of rejection in Group 2 showed vascular phase at 3 days, alveolar phase at 5 days, and destructive phase at 7 days, respectively. No evidence of rejection was found in Group 1 or 3. In Group 2, W3/25 positive cell proportion in GIL and BALF significantly decreased as the rejection progressed, but OX8 positive and OX39 positive cell proportion increases were significantly greater than in Groups 1 and 3 as the rejection progressed. These results lead us to speculate that the studies of T cell subsets in GIL and BALF lymphocytes are useful for diagnosis of rejection in lung transplantation.
Animals↗
Immunologic monitoring during OKT3 therapy.
The monoclonal antibody OKT3 is a potent immunosuppressant inducing both T-cell depletion and antigenic modulation of the CD3/T-cell receptor complex. CD3 + lymphocytes are cleared from the circulation within 1 hour of intravenous administration of OKT3. Periodic monitoring of remaining or reappearing CD3 + lymphocytes gauges the adequacy of OKT3 dosing and OKT3 consumption by anti-OKT3 antibodies. The maximum acceptable concentration of CD3 + lymphocytes for maintaining the effectiveness of OKT3 therapy remains controversial due to variable sensitivity of the methods used. Intravenously administered OKT3 has a half-life of approximately 18 hours and results in trough serum levels ranging from 500 to 1000 ng/ml in 3 to 4 days. The antibody response to OKT3 is oligoclonal, and both IgM and IgG antibodies to OKT3 may develop, showing restricted specificity; anti-isotypic and/or anti-idiotypic antibodies can be detected. The IgG anti-idiotypic antibodies are the ones that neutralize the therapeutic activity of OKT3, and their specific detection involves the use of an immunofluorescence method that tests their ability to block the binding of OKT3 to normal T cells. The production of OKT3 antibodies has been shown to be affected by concomitantly administered immunosuppressive agents. Studies have demonstrated that OKT3 can be successfully reused in allograft recipients who do not present anti-idiotypic blocking antibodies. The acute clinical syndrome regularly observed upon the first OKT3 injection is related to massive, although transient, release of several cytokines, which may be easily monitored with currently available radioimmunologic or immunoenzymatic tests.
Cytokines↗
Immunological monitoring of chronic pleural empyema patients.
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Adult↗
[Development of a rapid method of immunologic monitoring during pregnancy].
The authors validate the efficiency of enzyme-linked immunoassay (ELISA) for rapid diagnosis of rabies and titration of specific antibodies and of the method of isolation of field rabies strains in the rat Gasser's node neurinoma cells. Highly active specific diagnostic agents for ELISA and immunofluorescent test based on sheep immunoglobulins have been created for detection of the rabies agent in pathological material from different animals and humans.
Animals↗
[Clinico-immunologic monitoring of patients with gastroesophageal reflux during recuperative treatment].
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B-Lymphocytes↗
[Immunological monitoring of bone tissue resorption following endoprosthesis of large joints].
There was noted a reaction of the immune system to big-joint replacements, which developed both as a stereotype acute-phase response to the surgical intervention and a moderate immunosuppression, and as a response to the functioning of the implant (beginning from month 6 after surgery), when a mild phagocytes' activity and moderate changes in the acute-phase proteins were registered. Later, the monocyte-lymphocyte reaction of blood involving a higher production of osteoresorptive IL-1 and TNF were observed during the shaping resorption of osseous tissue. Complications can be prognosticated on the basis of the below: contents of leukocytes' main populations; activity of cytoplasm cation proteins; and concentration of the main classes of immunoglobulins and of IL-1.
Acute-Phase Proteins↗
[Immunological monitoring of sepsis using flow cytometry--quantitation of monocyte HLA-DR expression and granulocyte CD64 expression].
BACKGROUND: Sepsis is a serious disease with a high case fatality rate. A variety of changes in the host immune responsiveness are observed in the pathogenesis of sepsis, ranging from detrimental hyperinflammation to profound immunoparalysis, i.e. acquired immunodeficiency. The level of monocyte HLA-DR expression reflects the functional status of monocytes as antigen-presenting cells and granulocyte CD64 expression is also indicative of infectious inflammation. MATERIAL AND METHODS: Monocyte HLA-DR expression and granulocyte CD64 expression were measured in 49 septic patients and 30 healthy controls using flow cytometry focused on three parameters: positive cell percentage, mean fluorescence intensity and quantitation of antibodies bound per cell (QuantiBRITE). RESULTS: The significance of both monocyte HLA-DR expression and granulocyte CD64 expression in septic patients was confirmed. Monocyte HLA-DR dramatically decreases in septic patients compared to controls, is one of the prognostic factors and correlates with C-reactive protein. In contrast, granulocyte CD64 sharply rises in patients with sepsis and correlates with mediators of systemic inflammation (procalcitonin - PCT), proinflammatory mediators (interleukin-6 - IL-6, lipopolysaccharide binding protein - LBP) and anti-inflammatory cytokines (interleukin-10 - IL10). CONCLUSION: Quantitative monocyte HLA-DR expression and granulocyte CD64 expression are useful indicators in septic patients when considered along with the panel of other markers, monitored over a period of time and in the context of the clinical course of sepsis.
Adult↗