Lymphocyte subtypes CD3+, CD19+, CD16+CD56+, CD4+, CD8+, and CD3+HLA-DR+ in peripheral blood obtained from patients after thoracic organ transplantation

Introduction

The aim of this study was to assess peripheral blood lymphocyte subtypes (CD3⁺, CD19⁺, CD16⁺CD56⁺, CD4⁺, CD8⁺, and CD3⁺HLA-DR⁺) obtained from thoracic organ recipients at various periods after transplantation.

Material and Methods

Seventeen patients after lung transplantation (LT) and 5 patients after heart transplantation (HT) included 13 males (76.5%) and 4 females (23.5%) of overall mean age at the time of transplantation of 46.7 ± 11.55 years and mean body mass index of 21.1 ± 4. Lymphocyte phenotypes were estimated using Simultest IMK Plus.

Results

A significant decrease in lymphocytes of the majority of subtypes was observed at 1 year posttransplantation compared with normal ranges: CD19⁺ B lymphocytes in 56% of patients, CD8⁺ T cells among 48% and CD16⁺CD56⁺ natural killer elements, 56%. In contrast, there were increased numbers of activated lymphocytes (CD3⁺HLA-DR⁺). Beyond the 1-year observation, we observed a trend to normalize parameters among the majority of subjects.

Conclusion

A clear tendency to a decrease number of peripheral blood lymphocytes of various subtypes was observed among thoracic organ recipients in the first year posttransplantation with the exception of activated HLA-DR⁺ cells. After the first year, there was slow restoration of lymphocytes.     

Permanent pacing following cardiac transplantation.

Permanent pacemakers were inserted in 20 of 439 patients who had received 453 orthotopic cardiac allografts since 1980 at the Columbia-Presbyterian Medical Center. Mean age at transplantation was 45 +/- 4 (SEM) years (range 10 to 64). Pacemakers were inserted an average of 2.4 +/- 1 months after transplantation (range 0.4 to 29), 16 of 20 (80%) within the first month. Indications included sinus bradycardia or sinus arrest in 15 (75%), third-degree heart block in 2 (10%), and both sinus node and atrioventricular node dysfunction in 3 (15%). Rejection episodes and pacemaker insertion were associated in 8 patients (40%). Pacing modes included DDD (7 patients, 35%), AAI,R (7 patients, 35%), VVI,R (3 patients, 15%), DDD,R (2 patients, 10%), and VVI (1 patient, 5%). There was no pacing-related morbidity or mortality. Fourteen of 20 patients (70%) are alive and well 3 to 48 months (mean 24 +/- 4) after transplantation. Late follow-up indicated that atrioventricular node dysfunction resolved in one of two patients, sinoatrial node dysfunction improved or resolved in 7/13 patients, and no atrioventricular block developed in 11 (8 to 37 months, mean 22 +/- 3). Permanent pacing can be safely performed following orthotopic cardiac transplantation, predominantly for sinus node dysfunction. The requirement for pacing may reflect ongoing or new onset rejection and patients should therefore be evaluated accordingly. Dual-chamber pacing is probably not necessary unless atrioventricular node dysfunction is coexistent. Further, as most transplant recipients return to an active life-style, AAI,R may be the preferred mode of pacing.     

Psychiatric disorders and outcome following cardiac transplantation.

BACKGROUND: Psychosocial factors are frequently included in determining candidacy for cardiac transplantation. Although there are some data demonstrating a link between preoperative psychosocial status and postoperative outcome, a definitive answer has yet to be reached. METHODS: We studied 107 consecutive patients, transplanted from January 1990 to September 1991, with a retrospective review of pretransplant psychiatric evaluations to define a DSM III-R Axis I diagnosis. The medical outcome data included 1-year survival, rehospitalizations, infections, and rejection episodes, gathered from the transplant database. RESULTS: There were no discernable differences between the groups with (n = 25) and without (n = 82) a DSM III-R Axis I psychiatric disorder prior to transplant in the evaluation of demographic data and medical outcome variables. CONCLUSION: These data demonstrate that individuals with a history of psychiatric disorder who are carefully selected for compliance with medical care display no inherent difference from individuals without psychiatric disorder in medical outcome and survival at 1 year after cardiac transplantation.     

Decreased CD3+CD16+ natural killer-like T-cell percentage and zeta-chain expression accompany chronic inflammation in haemodialysis patients

Aim:   Clinical and experimental data indicate a deficient immune response in haemodialysis (HD) patients. Natural killer-like (NKL) T cells express on their surface both the T-cell antigen receptor (TCR) and a diverse set of NK-cell receptors (NKR) and share properties of both T cells and NK cells. ζ-Chain phosphorylation is an early event that follows TCR activation or some NKR activation. The ζ-chain of both T cell and NK cells is downregulated in many chronic inflammatory states, HD included. In the present study, NKL T-cell percentage and ζ-chain expression in HD patients were evaluated.
Methods:   Thirty-three stable HD patients and 30 healthy volunteers were enrolled into the study. NKL T-cell percentage and NKL T-cell ζ-chain mean fluorescence intensity (MFI) were evaluated with flow cytometry. The inflammatory markers C-reactive protein, interleukin-6 and tumour necrosis factor-α were measured in the serum by means of enzyme-linked immunosorbent assay.
Results:   All the evaluated markers of inflammation were increased in HD patients. In these patients, NKL T-cell percentage (1.71 ± 1.69% vs 3.94 ± 3.86%) and ζ-chain MFI (3.66 ± 2.79 vs 7.03 ± 7.91) were decreased.
Conclusions:   NKL T-cell percentage and ζ-chain expression is decreased in HD patients. Taking into consideration the continuously increasing age of the HD patients and that normally NKL T-cell numbers increase with age counteracting the impaired T-cell and NK-cell function accompanying advancing age, the above NKL T-cell disturbances could contribute to the impaired immune response in this population. Measures towards alleviating chronic inflammation could partially restore NKL T-cell impairment.     

T-cell activation, proliferation, and memory after cardiac transplantation in vivo

OBJECTIVE: To study the response of alloantigen (H2Kb)-specific T cells to a H2b+ cardiac allograft in vivo. SUMMARY BACKGROUND DATA: The response of T cells to alloantigen has been well characterized in vitro but has proved more difficult to assess in vivo. The aim of these experiments was to develop a model of T-cell-mediated rejection where the response of T cells after transplantation of a cardiac allograft could be followed in vivo. METHODS: Purified CD8+ T cells from H2Kb-specific TCR transgenic mice (BM3; H2k) were adoptively transferred into thymectomized, T-cell-depleted CBA/Ca (H2k) mice. These mice were then transplanted with a H2Kb+ cardiac allograft. Using four-color flow cytometry, the proliferative response, modulation of activation markers, and potential cytokine production of the H2Kb-specific T cells was assessed after transplantation. RESULTS: Consistent rejection of H2Kb+ cardiac allografts required the transfer of at least 6 x 10(6) CD8+ H2Kb-specific T cells. Short-term analyses revealed that the transgenic-TCR+/ CD8+ T cells proliferated and became activated after transplantation of an H2Kb+ cardiac allograft. Fifty days after transplantation, the transgenic-TCR+/CD8+ T cells remained readily detectable, bore a predominantly memory phenotype (CD44hi), and rapidly produced interleukin 2 and interferon-gamma on in vitro restimulation. CONCLUSIONS: These data show that the activation of alloantigen-specific T cells can be followed in vivo in short-term and long-term experiments, thereby providing a unique opportunity to study the mechanisms by which T cells respond to allografts in vivo.     

CMV-specific CD8 T-cell dynamics in the blood and the lung allograft reflect viral reactivation following lung transplantation.

Despite the potentially high burden of cytomegalovirus (CMV)-related disease following lung transplantation, the role of the cytotoxic T-lymphocyte (CTL) response to CMV in this patient group is ill-defined. We assessed the CMV-specific T-cell response in the blood and lung allograft of immunosuppressed lung transplant recipients receiving antiviral prophylaxis and following their withdrawal. While the proportion of CMV-specific CTL varied between patients, in the absence of CMV reactivation the level of CMV-specific CD8+ T cells in the blood remained stable over time. In the majority of patients CMV-specific cells could be detected in the lung allograft, often in the absence of viral DNA. Additionally, following primary CMV lung infection, CMV-specific CD8+ T cells were detected no earlier than 100 days post-transplantation but still prior to the detection of viral DNA in the lung allograft. Together these findings suggest that very low levels of CMV replication are sufficient to both prime and recruit CMV-specific CD8+ T cells to the MHC-mismatched lung allograft. The direct detection of CMV-specific T cells with an effector phenotype in the lung allograft suggests a protective antiviral function. This study provides a framework upon which the association between CMV and chronic allograft rejection can be further studied.     

Nodal involvement by cutaneous CD30-positive T-cell lymphoma mimicking classical Hodgkin lymphoma

An association between classical Hodgkin lymphoma (cHL) and mycosis fungoides (MF) or lymphomatoid papulosis has been reported in the literature. However, there can be considerable morphologic and immunophenotypic overlap between cHL and nodal involvement by CD30-positive T-cell lymphoproliferative disorders (CD30-T-LPD). To examine this potential association, biopsies from patients with a history of MF or primary cutaneous CD30-T-LPD and lymph node biopsies reported as either CD30-positive T-cell lymphoma (TCL) with Hodgkin-like cells or cHL were retrieved from the authors' institution. Of 11 cases identified, 10 were considered CD30-positive TCL with Hodgkin-like cells, whereas 1 was confirmed as cHL upon review. Five cases originally diagnosed as cHL were revised as CD30-positive TCL. Cases of CD30-positive TCL with Hodgkin-like cells showed a male predominance (M:F, 4:1) with a median age of 53 years (range, 44 to 72 y). Nearly all patients (9/10) initially presented with skin lesions. In 7/10 patients the draining lymph node was involved, whereas in 3 cases this could not be confirmed. Tumor cells morphologically resembled Hodgkin/Reed-Sternberg cells; they were uniformly strongly positive for CD30, and CD15 was expressed in 9/10 (90%) cases. A T-cell derivation was confirmed by T-cell antigen expression (7/10) and clonal rearrangement of T-cell receptor genes (9/10). In 3 cases a common T-cell clone was identified in skin and lymph node. B-cell markers (CD20/PAX5) were consistently negative. In 1 case the diagnosis of cHL followed by lymphomatoid papulosis was confirmed, with Hodgkin/Reed-Sternberg cells expressing PAX5, CD30, and CD15. In situ hybridization studies for Epstein Barr virus were negative. We show that cHL is less often associated with MF and primary cutaneous CD30-T-LPD than previously thought and that the coexpression of CD30 and CD15 in these TCLs may lead to a mistaken diagnosis of cHL.     

Expression of programmed death-1 in primary cutaneous CD4-positive small/medium-sized pleomorphic T-cell lymphoma, cutaneous pseudo-T-cell lymphoma, and other types of cutaneous T-cell lymphoma

In this study we investigated whether programmed death-1 (PD-1) could serve as a useful diagnostic marker to differentiate between primary cutaneous CD4 small/medium-sized pleomorphic T-cell lymphoma (PCSM-TCL) and cutaneous pseudo-T-cell lymphomas on the one hand and other types of cutaneous T-cell lymphomas (CTCLs) on the other. Formalin-fixed, paraffin-embedded skin biopsies from 26 patients with PCSM-TCL or pseudo-T-cell lymphoma, including 1 patient with a lymphomatoid drug eruption, and 52 skin biopsies from other types of CTCLs were stained for PD-1. In addition, PD-1-positive cases were stained with antibodies against BCL6, CXCL13, and CD10 to determine a possible relationship with follicular helper T (TFH) cells. In all 26 cases of PCSM-TCL or pseudo-T-cell lymphoma, the medium-sized to large-sized atypical T cells consistently expressed PD-1, BCL6, and CXCL13 but not CD10. PD-1 expression was found in only 2 of 21 cases of mycosis fungoides and in only 2 of 16 cases of cutaneous peripheral T-cell lymphoma, unspecified. All 4 patients with an aggressive epidermotropic cytotoxic CD8 CTCL and all 11 cases with a primary cutaneous CD30 lymphoproliferative disorder were negative for PD-1. In conclusion, PD-1 is typically expressed by atypical cells in PCSM-TCL and pseudo-T-cell lymphoma but is not expressed or is rarely expressed in other types of CTCLs. Therefore, it may serve as a suitable adjunct in differential diagnosis. Our results demonstrate that the atypical cells in PCSM-TCL and pseudo-T-cell lymphomas share a common TFH phenotype and support the view that most cases classified nowadays as PCSM-TCL are identical to cutaneous pseudo-T-cell lymphomas described previously.     

CD10 expression in extranodal dissemination of angioimmunoblastic T-cell lymphoma

Angioimmunoblastic T-cell lymphoma (AITL) is a systemic disease that often has evidence of extranodal involvement at presentation. In a recent study of lymph nodes in AITL, we showed that the neoplastic T cells in most cases can be identified by aberrant expression of CD10. The aim of this study was to investigate whether CD10 expression by the neoplastic T cells is maintained in extranodal sites. Ten cases of AITL with histologic and immunophenotypic evidence of extranodal dissemination were studied. Seven cases of peripheral T-cell lymphoma unspecified (PTLu), that included biopsies of involved extranodal sites, two cases of enteropathy type T-cell lymphoma (ETTL), and one case of extranodal NK/T lymphoma, nasal type were selected as controls. Diagnostic lymph node biopsies and biopsies of extranodal sites were reviewed. PCR for T-cell clonality and single layer immunostaining for CD3, CD20, CD10, and CD21 and double layer immunostaining for CD20/CD10 were performed. All 10 cases of AITL had characteristic histologic features and molecular evidence of the disease in lymph node biopsies. In these cases, aberrant CD10 expression was maintained in the lung, cecum, tonsil, nasopharynx, and one of six involved bone marrow trephines. In these extranodal biopsies, the distribution of CD10-positive tumor cells correlated with that of the follicular dendritic cell meshwork (FDC). The five bone marrow trephines that lacked aberrant CD10 expression were devoid of morphologic and immunohistochemical evidence of FDC. In these five cases, there was evidence of aberrant CD10 expression in other involved sites that had FDC. The neoplastic cells in PTLu, ETTL, and extranodal NK/T lymphoma, nasal type were CD10 negative. Our data show that aberrant CD10 expression is a useful phenotypic marker for diagnosis of AITL in most involved extranodal sites, except bone marrow, and suggest a possible role of FDC in the pathogenesis of AITL.     

Morbidity and mortality in diabetic patients following cardiac transplantation.

BACKGROUND: Diabetes remains a relative contraindication to cardiac transplantation. Previous reports have described small numbers of diabetic patients without end-organ damage who have undergone successful cardiac transplantation. METHODS: A retrospective analysis of diabetic patients transplanted and their outcome in a single large center from 1/1/95 to 12/31/99 was performed. Diabetes was defined as "medium risk" by the presence of any of the following parameters: duration of therapy >10 years; use of insulin; serum creatinine >2 mg/dl; urinary protein >300 mg per 24 hours; presence of peripheral vascular disease (ankle:brachial ratio <1.0); and documentation of other diabetic comorbidity (retinopathy, neuropathy, gastroparesis). RESULTS: During this time period, 374 adult cardiac transplants were performed. Seventy-six patients (20%) were diabetic with 33 patients (43%) requiring insulin. Forty-two of the patients had moderate disease. Survival of the diabetic and non-diabetic recipients was comparable (1- and 3-year survival of 86% and 85%. vs 87% and 84%, respectively, p = NS). No difference in survival between "medium-risk" and "low-risk" diabetics was observed. The incidence of acute rejection in the first year, graft vasculopathy and infection, was comparable between diabetic and non-diabetic patients. In both diabetic and non-diabetic patients, there was a similar and small insignificant increase in serum creatinine. CONCLUSIONS: More patients with advanced diabetes are undergoing cardiac transplantation and the early and mid-term survival remains comparable to non-diabetic recipients. Future liberalization of transplantation in diabetics appears likely.     

CD8+ T-cell maturation following lung transplantation: the differential impact of CMV and acute rejection

Studies on persistent viral infections demonstrate that CD8(+) T-cells differentiate along distinct pathways following chronic antigen exposure; however the effect of stimulation with non-viral chronic antigens is poorly described. We assessed the contributions that the presence of an allograft or cytomegalovirus (CMV) has on the post-thymic differentiation of CD8(+) T-cells in both the blood and lung allograft in patients undergoing lung transplantation. CD28 expression on blood CD8(+) T-cells was reduced in CMV seropositive patients, and was further reduced following acute episodes of CMV reactivation. These viral-associated changes in phenotype were not seen in CD8(+) T-cells isolated from the lung allograft where a different pattern of CD28 expression was observed. Following transplantation there was a progressive reduction in CD28 expression on BAL CD8(+) T-cells. In contrast to what was observed in peripheral blood, reduced CD28 expression on BAL CD8(+) T-cells was associated with a reduced gamma-IFN production. Furthermore, a high proportion of CD28-CD8(+) T-cells in the BAL was associated with fewer episodes of acute allograft rejection. An expanded CD28-CD8(+) T-cell subset, with reduced function, within the lung allograft may have important prognostic implications in lung transplant recipients.     

Mechanical assistance for cardiogenic shock following cardiac surgery, myocardial infarction, and cardiac transplantation

From January, 1982, to October, 1986, 33 patients were treated with either the Pierce-Donachy prosthetic ventricle or the Bio-Medicus ventricular assist device for cardiogenic shock following a cardiac operation, myocardial infarction, or cardiac transplantation. Twenty-five patients required the assistance for postcardiotomy shock and 8, for a variety of conditions including myocardial infarction shock and myocarditis, and as a bridge to cardiac transplantation. Complications were frequent and usually secondary to prolonged cardiopulmonary bypass. Results were poorest in the group with postcardiotomy shock. Earlier application of an assist device could lead to more frequent survival and avoidance of the detrimental effects of prolonged extracorporeal circulation.     

Primary cutaneous CD4+ small/medium-sized pleomorphic T-cell lymphoma expresses follicular T-cell markers

Cutaneous CD4 small/medium-sized pleomorphic T-cell lymphoma (CSTCL) is a cutaneous T-cell lymphoma defined by a predominance of small-to-medium-sized CD4 pleomorphic T cells, with a favorable clinical course. Cases are also characterized by the presence of a rich infiltrate of reactive B cells. Recently, it has been reported that follicular helper T cells (TFH cells) display a distinct gene expression profile, positive for PD-1, CXCL13, and BCL-6. We report for the first time the expression of PD-1 and other TFH cell markers in CSTCLs and discuss its biologic significance. Sixteen CSTCLs were included in this study, and also 20 reactive inflammatory conditions, 10 primary cutaneous marginal zone, 10 follicular center lymphomas, and 5 primary CD30 cutaneous lymphomas. They were immunohistochemically analyzed for a large panel of markers. Double immunoperoxidase labeling of paraffin sections was performed for PD-1, OCT-2, and BCL-6. Clonal Ig and T-cell receptor rearrangements and Epstein-Barr virus-encoded RNA expression were also evaluated. Morphologic and clinical data were reviewed. Histologic examination showed a dense polymorphic lymphoid infiltrate throughout the dermis. Atypical large CD4 cells were positive for PD-1, CXCL13, and BCL-6 in all cases, and were attached in small clusters, or formed rosettes around CD30/OCT-2+ B blast cells. Epstein-Barr virus was not apparent in any of the cases. A dominant T-cell clone was identified in 14 cases, whereas polymerase chain reaction IgH gene rearrangement studies showed that all cases were polyclonal. None of the patients had lymphadenopathy or showed any evidence of systemic disease, nor did they have any previous history of mycosis fungoides or drug reactions. FTH cell markers are not exclusive to angioimmunoblastic lymphadenopathy but may also be seen in neoplastic cells of CSTCLs. Moreover, these findings suggest that B-cell stimulation by FTH could also take place in some cutaneous T-cell lymphomas.