scholarly journals Nodular Lymphocyte Predominant Hodgkin Lymphoma - A Retrospective Immunohistochemical Study of Patients in Bangalore, Karnataka

2021 ◽  
Vol 8 (23) ◽  
pp. 1966-1969
Author(s):  
Shankar Anand ◽  
Akshatha C ◽  
Libin Babu Cherian ◽  
Ramachandra C
Keyword(s):  
T Cells ◽  
T Cell ◽  
Lymph Nodes ◽  
Hodgkin Lymphoma ◽  
Classical Hodgkin Lymphoma ◽  
Double Positive ◽  
Cervical Lymph Nodes ◽  
Histiocytic Cell ◽  
Immunohistochemical Markers ◽  
Lp Cells

BACKGROUND The term Hodgkin’s lymphoma includes classical Hodgkin lymphoma (CHL) and the less common nodular lymphocyte predominant Hodgkin lymphoma (NLPHL). NLPHL is a B cell neoplasm usually characterised by nodular or follicular and diffuse proliferation of small lymphocytes with single scattered large neoplastic cells (LP/L&H/Popcorn cells). NLPHL accounts for 10 % of all Hodgkin lymphoma. METHODS This is a retrospective study. Histopathology slides and blocks of 24 cases of nodular lymphocyte predominant Hodgkin lymphoma were collected from the archives of histopathology from 2011 to 2015. The immunohistochemistry slides of the corresponding histopathology cases were also assembled. Both the slides were reviewed by three expert onco-pathologists and IHC markers were studied and compared. RESULTS Patients were mostly young between 20 and 40 years (16 / 24, 66.67 %). There was a distinct male preponderance (20 / 24, 83.3 %). Most cases involved cervical, axillary or inguinal lymph nodes, with cervical lymph nodes being the most common (13 / 24, 54 %). It was found that CD45, CD20, CD79a and PAX5 staining highlighted the LP cells in all twenty-four cases, while OCT - 2 and BOB - 1 were highlighted in twenty-three cases (95.8 %), which was statistically significant. CD3 and CD5 IHC staining on T cell rosettes and background reactive T cells were examined, and it was seen that CD3 expression was far more consistent than CD5 expression in T cell rosettes and reactive T cells. Also, it was seen that, those cases which were double positive for CD3 and CD5 constitutes only eight cases (8 / 24, 33.3 %). CONCLUSIONS CD3 is a more consistent marker than CD5 in demonstrating surrounding reactive T cells in NLPHL. CD45, PAX5, CD20, BOB - 1 and OCT - 2 are consistent immunohistochemical markers of LP cells. KEYWORDS Nodular Lymphocyte Predominant Hodgkin Lymphoma (NLPHL), Classical Hodgkin Lymphoma (CHL), Cluster Differentiation (CD), Lymphocyte Predominant Cells (LP Cells), Lymphocyte and Histiocytic Cell (L & H Cell)

Identification and Purification of Hodgkin Cells from Lymph Nodes Involved by Classical Hodgkin Lymphoma by Flow Cytometry and Flow Cytometric Cell Sorting.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 2262-2262
Author(s):  
Jonathan R. Fromm ◽  
Steven J. Kussick ◽  
Brent L. Wood
Keyword(s):  
Flow Cytometry ◽  
T Cells ◽  
T Cell ◽  
Lymph Nodes ◽  
Hodgkin Lymphoma ◽  
Cell Sorting ◽  
Classical Hodgkin Lymphoma ◽  
Tissue Sections ◽  
Hla Dr ◽  
Blocking Antibodies

Abstract The diagnosis of classical Hodgkin lymphoma (CHL) has historically been made in tissue sections, as attempts to identify the neoplastic Hodgkin and Reed-Sternberg (HRS) cells of CHL by flow cytometry (FC) have been largely unsuccessful. As HRS cells are known to be ringed (“rosetted”) by benign/reactive T cells, we hypothesized that in cell suspensions the HRS will be bound to T cells (forming T cell rosettes), and that consequently the rosettes would have a composite T-cell/HRS immunophenotype by FC (CD3+/CD15+/CD20−/CD30+/CD45+). We further hypothesized that specific antibodies to the adhesion molecules known to be involved in T cell/HRS cell binding (CD2 and LFA-1 on the T cell, and CD54 and CD58 on the HRS cell) might result in “naked” (unbound) HRS cells, enabling us to use FC to identify HRS cells with the expected immunophenotype (CD3−/CD15+/CD20−/CD30+/CD45−). Our initial FC studies of the HRS cell line L1236 demonstrated that CD15, CD30, CD40, CD71, CD86, CD95, and HLA-DR, but not CD3 or CD20, were brightly expressed on these cells and may be useful in identification of HRS in authentic cases of CHL involving lymph nodes. In mixing experiments, L1236 cells spontaneously bound normal T cells, analogous to T cell rosetting of HRS cells in CHL; these interactions could be blocked specifically using a cocktail of unlabeled antibodies to CD2, LFA-1, CD54, and CD58. Among 27 lymph nodes involved by CHL, this novel FC method, in which 250,000 to 500,000 total lymph node cells were evaluated, and in which up to ten cellular antigens were assessed simultaneously, enabled HRS cells to be identified in 89% of cases. 82% of these cases demonstrated interactions between HRS cells and T cells that could be disrupted with blocking antibodies. None of 29 non-CHL neoplasms, and none of 23 reactive lymph nodes, demonstrated HRS populations by FC. The proportions of cases where the HRS population showed expression of CD15, CD30, CD40, CD71, CD86, CD95, and HLA-DR, and absence of CD3 and CD20 was similar to that described previously in tissue sections by immunohistochemistry. Interestingly, in contrast to the findings in tissue sections, by FC the non-rosetted HRS cells of most CHL cases (73%) demonstrated detectable expression of CD45, usually at a low level. Finally, cell sorting experiments confirmed that (1) populations identified by FC have the cytomorphology of HRS cells, (2) HRS/T cell rosettes can be detected by FC, and (3) these rosettes can disrupted by the blocking antibody cocktail. This FC technique offers a potential alternative to immunohistochemistry in confirming the diagnosis of CHL and, through cell sorting, provides a means of rapidly purifying HRS cells.

CD4+CD26 - T Cell Population in Classical Hodgkin Lymphoma Displays a Distinctive Regulatory T Cell Population.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 384-384
Author(s):  
Yue Ma ◽  
Lydia Visser ◽  
Tjasso Blokzijl ◽  
Geert Harms ◽  
Cigdem Atayar ◽  
...  
Keyword(s):  
T Cells ◽  
T Cell ◽  
Lymph Nodes ◽  
Hodgkin Lymphoma ◽  
Regulatory T Cell ◽  
T Cell Subsets ◽  
Mrna Levels ◽  
Classical Hodgkin Lymphoma ◽  
Cell Type ◽  
Activated T Cell

Abstract Introduction: Hodgkin and Reed-Sternberg (HRS) cells in classical Hodgkin lymphoma (cHL) are surrounded by a majority of infiltrating reactive cells, which mainly consists of CD4+ T cells. These T cells express several activation associated surface markers but lack expression of the T cell co-stimulatory molecule CD26. Little is known about the significance of these rosetting CD4+CD26- T cells. Methods: To characterize these T cells, CD4+CD26- and CD4+CD26+ T cells were sorted from lymph node cell suspensions from 7 cHL and 5 reactive lymph nodes (LN). Of 5 HL cases and 3 lymph nodes, parts of the cells were stimulated with PMA/ionomycin to get activated T cell subsets. mRNA profiles of activated and non-activated T cell populations were evaluated with quantitative RT-PCR for 46 selected genes. Results: We observed a higher percentage of CD4+CD26- T cells in cHL compared to reactive LN. For the non-activated T cell subsets, CD4+CD26- T cells in cHL showed higher mRNA levels of IL2RA, CTLA4, TNFRSF4 and CCR4 compared to LN. Moreover, these cells displayed low or no expression of the Th1 or Th2 related cytokines IL2, IFNγ, IL13, IL12B, IL4, IL5 and the chemoattractant receptor GPR44. Overall, the profiling results support a regulatory T (Treg) cell type for the CD4+CD26- T cells in cHL. Besides Tregs, Th17 cells may exist in cHL based on the significantly higher IL17 mRNA level for both the CD26- and CD26+ T cells in cHL than in LN. Upon activation, the lack of up-regulation of mRNA levels of most cytokine genes (IFNγ, IL2, IL8, IL21, IL17, IL13, IL12A and IL4) indicated an anergic character for the CD4+CD26− subset in cHL. Conclusion: A high proportion of CD4+CD26− T cells is characteristic for cHL. No evidence for a Th1 or Th2 cell type is found for these cells but they display a regulatory T cell phenotype. Anergy fits with the regulatory T cell profile of these cells, probably explaining the immunosuppressive mechanism involved in cHL.

CD4+ T cells are essential for the development of destructive thyroiditis induced by anti–PD-1 antibody in thyroglobulin-immunized mice

2021 ◽  
Vol 13 (593) ◽  
pp. eabb7495
Author(s):  
Yoshinori Yasuda ◽  
Shintaro Iwama ◽  
Daisuke Sugiyama ◽  
Takayuki Okuji ◽  
Tomoko Kobayashi ◽  
...  
Keyword(s):  
T Cells ◽  
T Cell ◽  
Lymph Nodes ◽  
Mononuclear Cells ◽  
Critical Role ◽  
T Cell Subsets ◽  
Effector Memory ◽  
Histopathological Analysis ◽  
Activated T Cells ◽  
Cervical Lymph Nodes

Immune-related adverse events induced by anti–programmed cell death–1 antibodies (PD-1-Ab), including destructive thyroiditis (thyroid-irAE), are thought to be caused by activated T cells. However, the T cell subsets that are directly responsible for damaging self-organs remain unclear. To clarify which T cell subsets are involved in the development of thyroid-irAE, a mouse model of thyroid-irAE was analyzed. PD-1-Ab administration 2.5 months after immunization with thyroglobulin caused destructive thyroiditis. Thyroiditis was completely prevented by previous depletion of CD4+ T cells and partially prevented by depleting CD8+ T cells. The frequencies of central and effector memory CD4+ T cell subsets and the secretion of interferon-γ after stimulation with thyroglobulin were increased in the cervical lymph nodes of mice with thyroid-irAE compared with controls. Histopathological analysis revealed infiltration of CD4+ T cells expressing granzyme B in thyroid glands and major histocompatibility complex class II expression on thyrocytes in mice with thyroid-irAE. Adoptive transfer of CD4+ T cells from cervical lymph nodes in mice with thyroid-irAE caused destruction of thyroid follicular architecture in the irradiated recipient mice. Flow cytometric analyses showed that the frequencies of central and effector memory CD4+ T cells expressing the cytotoxic marker CD27 were higher in peripheral blood mononuclear cells collected from patients with thyroid-irAE induced by PD-1-Ab versus those without. These data suggest a critical role for cytotoxic memory CD4+ T cells activated by PD-1-Ab in the pathogenesis of thyroid-irAE.

Tumor Microenvironment In Nodular Lymphocyte Predominant Hodgkin Lymphoma (NLPHL) Influences Occurrence of Relapses and Progression to Large Cell Lymphoma

Blood ◽  
2010 ◽  
Vol 116 (21) ◽  
pp. 2684-2684
Author(s):  
Nasir Bakshi ◽  
Mansoor Aljabry ◽  
Saad Akhter ◽  
Irfan Maghfoor ◽  
Ayman Mashi
Keyword(s):  
T Cells ◽  
T Cell ◽  
B Cells ◽  
Tumor Microenvironment ◽  
B Cell ◽  
Hodgkin Lymphoma ◽  
Clinical Course ◽  
Cell Lymphoma ◽  
Lp Cells

Abstract Abstract 2684 NLPHL accounts for 6.5% of all Hodgkin lymphoma cases in the West. It is characterized by a nodular or a nodular & diffuse proliferation of scattered large atypical CD20+ neoplastic B-cells referred to as lymphocyte predominant (LP) cells and typically associated with small lymphocytes mainly of B-cell type. Patients with NLPHL typically have an indolent clinical course but can frequently relapse. Progression to a higher grade lymphoma, notably T-cell/Histiocyte rich B-cell lymphoma (T/HRBCL) has been described in a relatively small number of cases. Because of its rarity, limited information is available about the role of non-neoplastic lymphocytes in NLPHL. Some studies suggest that NLPHL with T-cell rich background may behave differently than the conventional type with predominance of B-cells within the nodules. The purpose of this study was to evaluate outcomes of differential tumor microenvironment namely B-cell versus T-cell rich in patients with NLPHL. We document the clinicopathologic profiles of 29 patients with biopsy proven NLPHL, consisting of 22 male & 7 female, median age 26 years (range, 13–80 years). All patients had lymphoadenopathy & 2 cases showed extranodal involvement in addition to nodal disease. Two patients had a bulky mass, and three had stage 4 disease at presentation. The pathological diagnoses was reviewed and confirmed by an expert hematopathologist in all 29 cases. The LP cells in all cases had a prototypic immunophenotype of CD20+, CD79a+, PU.1+, Bcl-6+, CD15− CD30− & Fascin−. T/HRBCL was excluded as all cases demonstrated preservation of follicular dendritic meshwork by CD21 staining. The meshwork was expanded in 20 cases & in 9 cases it was partially disrupted evincing an irregular architectural pattern. Epstein-Barr Virus encoded RNA by in situ hybridization was negative in 8/8 cases tested. 27/29 patients received systemic multi-agent chemotherapy consisting of: doxorubicin, bleomycin, vinblastine, and dacarbacin (ABVD), 24 patients; cyclophosphamide, doxorubicin, vincristin, and prednisone (CHOP), 2 patients; Rituximab + CHOP (R-CHOP), 1 patient. 9/29 (31%) cases underwent autologous stem cell transplant. One patient in stage 2A refused therapy and one patient (stage 3A) developed significantly decreased cardiac ejection fraction following initial 2 cycles of ABVD. Both of these cases did not have adequate follow-up information available. Results: Twelve of the 29 cases (42%) were designated as having T-cell rich background population, whereas 17 (58%) were considered as conventional variant with a vast predominance of non-neoplastic small lymphocytes being B-cells. A few of the cases seemed to show admixture of both B-cells & T-cells. Comparing T-cell rich & B-cell rich background NLPHL no significant differences were detected in clinical parameters: age, sex, and stage at presentation, absolute lymphocyte count, LDH & Hb. All 27 (100%) patients in this study responded to first-line treatment: 23 with complete response & 4 with partial response. 13/27 (48%) had relapse/s. Five cases had more than one relapses. No patient died within a clinical follow-up period ranging from 18 to 84 months. When the overall survival (OS) of T-cell rich NLPHL was compared with the conventional variant there was no statistical significance between the two groups (log rank p= 0.1206). However, comparison of relapse rate showed that cases with T-cell rich background had higher relapse rate as well as greater incidence of multiple relapses as compared to B-cell rich type of NLPHL even after adjusting for the type of treatment received (log rank p= 0.003). Moreover, 2/12 (17%) T-cell rich NLPHL cases showed transformation to a high grade lymphoma (both T/HRBCL) at the time of recurrence. These findings suggest that in NLPHL a tumor microenvironment rich in T-cells rather than B-cells is characterized by an unfavorable clinical course although OS appears to be similar. These cases perhaps represent a distinctive clinicopathologic variant within the framework of NLPHL. Lately, the term ‘NLPHL with nodules resembling T/HRBCL’ has been used to express the immunobiological overlap between these two entities. It is possible that such cases could be regarded as “intermediate lymphomas” treading between NLPHL and T/HRLBCL. Further studies using gene array profiling analysis may help clarify the molecular differences between these closely related entities. Disclosures: No relevant conflicts of interest to declare.

Continuous CD4+ T Cell Lines Derived From the Classical Hodgkin Lymphoma Microenvironment: A Challenge to the Assumption of Anergy,

Blood ◽  
2011 ◽  
Vol 118 (21) ◽  
pp. 3645-3645
Author(s):  
Paul Greaves ◽  
Sameena Iqbal ◽  
David C Taussig ◽  
John G. Gribben
Keyword(s):  
T Cells ◽  
T Cell ◽  
Hodgkin Lymphoma ◽  
Cell Lines ◽  
Culture Media ◽  
Tissue Bank ◽  
Lymphocyte Culture ◽  
Classical Hodgkin Lymphoma ◽  
T Cell Lines ◽  
Reactive Node

Abstract Abstract 3645 Introduction: The bulk of the tumour infiltrate in classical Hodgkin lymphoma (CHL) is composed of immune cells, predominantly CD4+ T cells, with the malignant Hodgkin Reed Sternberg cell (HRS) representing <1% of cells. The lymphoid microenvironment has been described as anergic and hypoproliferative with suppressive properties (Marshall et al. Blood 2004 103:1755–62) but the functional significance of this is unclear. This study set out to examine the proliferative capacity and phenotype of T cells derived from CHL-diagnostic lymph node tissue taken at diagnosis. Method: Frozen single cell suspensions (SCS) from 6 patients were selected from the tissue bank of our Institute. T cell growth-augmenting and/or Th2 polarising cytokines were added in various combinations (IL2, IL4 only, IL2+4 or no added cytokine) to SCS-derived cells in 96 well plates at 0.3 × 106 cells per well in 200mcl of optimized lymphocyte culture media. No CD4+ enrichment step was carried out: all recovered cells were plated at baseline to maintain potential interactions between CD4+ cells and other cells, and no mitogen or T-cell receptor-stimulating or costimulating agents were added at any point. As controls, SCS derived from normal tonsil, and ÔreactiveÕ lymph nodes (n=4) (confirmed by histological report at the time of diagnosis) were also plated. Plates were examined daily for cell/colony morphology to estimate growth and split with fresh media and cytokines once every 7 days, with estimated proliferation (by haemocytometry) plotted. Cultures were assessed at baseline, 10 days, 28 days, 50 days and 100 days. Results: Proliferation, based on formation of discrete colonies and blastoid cell morphology, occurred in the majority of wells by day 7 in all CHL-derived cultures, and in a minority of wells, and to a lesser extent in all control cultures. CHL-derived T cells from one patient continue to expand after 200 days, doubling every 3–5 days, while the other 5 continue after 50–100 days. In contrast, no tonsil or reactive node-derived T cells survived beyond 50 days and none showed a net expansion in cell numbers. Growth was superior in the IL2+4 and IL2-only conditions, with no growth in the media-only or IL4-only conditions. The most favorable condition was with the addition of IL2+4. By day 21 a net increase in CHL-derived T cells was apparent, but not in any control T cell populations (Figure). At baseline, composition of the CHL-derived cells revealed a majority of CD3+ cells as expected, of which 60–80% were CD4+ and the remainder CD8+. By D21 the CD4+ component had outgrown all other cells in the CHL-derived cultures, being CD3+CD4+CD45RO+ consistent with antigen-experienced T helper cells, while all tonsil and reactive node-derived cells were CD8+CDRO+. Markers of central memory (CCR7 & CD62-L), Th2 (CCR4 and IL4), Treg (FOXP3 and CD25) and anergy (CD57) were absent after expansion, while markers of activation were upregulated (CD28, CD27, CD69, CD40L, CD30 & CD95). This phenotype persisted in the ongoing T cell lines. Conclusions: The CD4+ compartment of the CHL microenvironment contains a primed subset of cells capable of massive expansion without further mitogenic stimulation and of generating cytokine-dependent continuous cell lines with an antigen-experienced, activated phenotype. This challenges the assumption of T cell anergy and hypoproliferation in the tissue microenvironment of CHL. We are currently assessing the function and anti-tumor specific or tumor supportive nature of these T cells. Disclosures: Gribben: Roche: Honoraria; Celgene: Honoraria; GSK: Honoraria; Mundipharma: Honoraria; Gilead: Honoraria; Pharmacyclics: Honoraria.

scholarly journals The Tumor Microenvironment of Nodular Lymphocyte Predominant Hodgkin Lymphoma Is a Unique Immunobiological Entity Distinct from Classical Hodgkin Lymphoma

Blood ◽  
2018 ◽  
Vol 132 (Supplement 1) ◽  
pp. 4123-4123
Author(s):  
Jay Gunawardana ◽  
Karolina Bednarska ◽  
Soi C Law ◽  
Justina Lee ◽  
Muhammed Bilal Sabdia ◽  
...  
Keyword(s):  
T Cells ◽  
T Cell ◽  
Board Of Directors ◽  
Hodgkin Lymphoma ◽  
Research Funding ◽  
Host Immunity ◽  
Immune Checkpoints ◽  
Classical Hodgkin Lymphoma ◽  
Advisory Committees ◽  
Immune Effector

Abstract There is proven pre-clinical and clinical efficacy of mono or combinatorial immune strategies to boost host anti-lymphoma immunity, with classical Hodgkin Lymphoma (cHL) seen as the 'poster child'. Approaches include blockade of immune-checkpoints on exhausted tumor-specific T-cells (via mAb blockade of PD-1, TIM3, LAG3, TIGIT or their ligands), activation of T-cells via mAbs agonistic to CD137, and finally modulation of FOXP3, CTLA-4 and/or LAG3 regulatory T-cells (Tregs) or immunosuppressive tumor-associated macrophages (TAMs). In contrast, studies characterizing the circulating and intra-tumoral microenvironment (TME) of the distinct but rare CD20+ Hodgkin Lymphoma entity (5-8% of HL), Nodular Lymphocyte Predominant Hodgkin Lymphoma (NLPHL), are minimal. Furthermore, to our knowledge no functional profiling studies comparing the host immunity of NLPHL with cHL has been performed. We compared host immunity in 29 NLPHL patients, 30 cHL patients and 10 healthy individuals, with a focus on pertinent and clinically actionable immune parameters. Paraffin-embedded tissue and paired (pre- and post-therapy) peripheral blood mononuclear cells samples were interrogated by digital multiplex hybridization (Nanostring Cancer Immune Profiling Panel) and flow cytometry. Although cytotoxic T-cell gene counts (CD8a, CD8b) were similar, compared to cHL there were higher levels of the immune effector activation marker CD137 (gene counts 439 vs. 287; P<0.01). Consistent with this, CD4 and the Treg markers LAG3, FOXP3 and CTLA-4 were lower in NLPHL (2-4 fold lower, all P<0.05), with no difference in T-helper cell activation markers CD40L and CD30L seen between tumors. TAMs and dendritic cell markers MARCO, CD36, CD68, CD163, COLEC12 and CD11b were all lower in NLPHL than cHL (all P<0.05). In line with the known 'rossette' formed around LP cells by PD-1+ T-lymphocytes, we observed strikingly elevated PD-1 and the other T-cell checkpoints TIM3 and TIGIT in NLPHL (all 2-3 fold, P<0.001). However, in line with the known gene amplification of PD-L1 on HRS cells and its presence on TAMs, gene counts of this checkpoint ligand were 2-fold higher in cHL (P<0.001). Flow cytometry profiling of immune subsets in peripheral blood showed findings consistent with findings in the TME. Specifically, there was elevation of multiple exhaustion markers within CD4, CD8, and NK immune effector cells, with a striking proportion of highly anergic dual-LAG3/PD-1 positive CD8+ T-cells. Also there was elevation of immune-suppressive monocyte/macrophages in cHL relative to NLPHL. Relative to healthy lymph nodes, there was prominent up-regulation of a range of T-cell associated exhaustion markers in both NLPHL and cHL, indicating dysregulated priming of effector immune responses and host immune homeostasis. Comparison between NLPHL and cHL illustrated that NLPHL had a myriad of features that marked its intratumoral TME as a unique immunobiological entity typified by elevated immune checkpoint markers and T-cells with a highly anergic phenotype. Put together, these findings indicate that distinct immune evasion mechanisms are operative within the TME of NLPHL, including markedly higher levels of multiple immune-checkpoints relative to cHL. In contrast, Treg subsets and immune-suppressive monocyte/macrophages were relatively lower than that seen in cHL. T-cells frequently had dual immune-checkpoint expression. The findings from this study provides a compelling pre-clinical rationale for targeting PD-1 or combinatory checkpoint inhibition in NLPHL and sets the basis for future 'chemo-free' rituximab + checkpoint inhibitor clinical trials. Disclosures Tobin: Amgen: Other: Educational Travel; Celgene: Research Funding. Birch:Medadvance: Equity Ownership. Keane:Takeda: Other: Educational Meeting; BMS: Research Funding; Roche: Other: Education Support, Speakers Bureau; Celgene: Consultancy, Research Funding; Merck: Consultancy. Gandhi:BMS: Membership on an entity's Board of Directors or advisory committees, Research Funding; Celgene: Membership on an entity's Board of Directors or advisory committees, Research Funding; Janssen: Honoraria, Membership on an entity's Board of Directors or advisory committees; Merck: Honoraria, Membership on an entity's Board of Directors or advisory committees; Amgen: Honoraria; Takeda: Honoraria; Gilead: Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding.

Impact of Treatment On CD4+CTLA-4+ T Cell Subset in Brazilian Patients with Classical Hodgkin Lymphoma

Blood ◽  
2012 ◽  
Vol 120 (21) ◽  
pp. 4778-4778
Author(s):  
Joyce M. K. Silva ◽  
Maria Mirtes Sales ◽  
Adriana M. Damasco Penna ◽  
Elma Maria Chaves Maria Chaves ◽  
Priscilla B Silva ◽  
...  
Keyword(s):  
Immune Response ◽  
T Cells ◽  
T Cell ◽  
T Lymphocytes ◽  
Hodgkin Lymphoma ◽  
Cd4 T Cells ◽  
Surface Expression ◽  
Post Treatment ◽  
Healthy Controls ◽  
Classical Hodgkin Lymphoma

Abstract Abstract 4778 Cytotoxic T lymphocyte antigen-4 (CTLA-4) is one of the basic antigens involved in immune responses regulation associated with autoimmune diseases and cancer. Its key role in regulating the immune system has made CTLA-4 an attractive target for cancer. Augmentation of the immune response via blockade of CTLA-4 has shown an improvement in survival for patients with metastatic melanoma, which prompted the Food and Drug Administration (FDA) approval of the CTLA-4 function blocking antibody Ipilimumab for this disease. Objective: The aim of the study was to evaluate the surface expression of CTLA-4 on CD4+ T cells in peripheral blood mononuclear cells (PBMC) of patients with classical Hodgkin lymphoma (cHL) at diagnosis and post-treatment and correlate these findings with clinical and epidemiological aspects. Material and Methods: This is an open study and, so far, we included 35 patients from December 2009 to December 2011. Blood was drawn at diagnosis and post-treatment (1 to 4 months after completion of therapy). The T cell phenotype was evaluated by flow cytometry using CD3, CD4, CD8, CTLA-4 and correlated to phenotypic and clinical parameters in uni- and multivariate models pre and post-treatment. Eighteen healthy blood donors volunteers were recruited as controls. In this study, only cHL patients whose histology could be confirmed and Epstein-Barr (EBV) association established were studied. All patients were HIV negative and received ABVD chemotherapy protocol and radiotherapy if necessary. Three patients relapsed, and blood was also drawn at this time. Results: From the 35 cHL patients, 17 were EBV related and 18 EBV non-related. The percentage of CD4+ T cells with CTLA-4 surface expression was significantly increased in patients with cHL at diagnosis compared with healthy controls (median 7.36 vs 2.73; P<0.001). Additionally, CD4+CTLA-4+ T lymphocytes significantly decreased following treatment and complete response (7.36 vs 4.53; p=0.008), with values similar to healthy controls (4.53 vs 2.73; p=0.07). Interestingly, CD4+CTLA-4+ T lymphocytes on relapse were significantly different from post-treatment values and similar to pre treatment. There was no difference on CD4+CTLA-4+ T lymphocytes in the EBV related and non-related cHL patients. Regarding patient's baseline characteristics, CD4+CTLA-4+ T lymphocytes strongly correlated with erythrocyte sedimentation rate (ESR) values (r=0.67; p=0.002). Conclusions: We showed that CD4+CTLA-4+ T lymphocytes are increased in Brazilian cHL patients at diagnosis compared with post-treatment values and healthy controls. These results suggest a role of CTLA-4 on Hodgkin lymphomagenesis, possibly negatively regulating host anti-tumor immune response. The promising immunotherapy regimen targeting CTLA-4 might be beneficial in classical Hodgkin lymphoma. Disclosures: No relevant conflicts of interest to declare.

scholarly journals In Vivo Activation of Naive CD4+ T Cells in Nasal Mucosa-Associated Lymphoid Tissue following Intranasal Immunization with Recombinant Streptococcus gordonii

2006 ◽  
Vol 74 (5) ◽  
pp. 2760-2766 ◽  
Author(s):  
Donata Medaglini ◽  
Annalisa Ciabattini ◽  
Anna Maria Cuppone ◽  
Caterina Costa ◽  
Susanna Ricci ◽  
...  
Keyword(s):  
T Cells ◽  
T Cell ◽  
Lymph Nodes ◽  
Nasal Mucosa ◽  
T Cell Activation ◽  
Lymphoid Tissue ◽  
Streptococcus Gordonii ◽  
Cervical Lymph Nodes ◽  
Intranasal Immunization

ABSTRACT The antigen-specific primary activation of CD4+ T cells was studied in vivo by adoptive transfer of ovalbumin-specific transgenic T cells (KJ1-26+ CD4+) following intranasal immunization with recombinant Streptococcus gordonii. A strain of S. gordonii expressing on its surface a model vaccine antigen fused to the ovalbumin (OVA) peptide from position 323 to 339 was constructed and used to study the OVA-specific T-cell activation in nasal mucosa-associated lymphoid tissue (NALT), lymph nodes, and spleens of mice immunized by the intranasal route. The recombinant strain, but not the wild type, activated the OVA-specific CD4+ T-cell population in the NALT (89% of KJ1-26+ CD4+ T cells) just 3 days following immunization. In the cervical lymph nodes and in the spleen, the percentage of proliferating cells was initially low, but it reached the peak of activation at day 5 (90%). This antigen-specific clonal expansion of KJ1-26+ CD4+ T cells after intranasal immunization was obtained with live and inactivated recombinant bacteria, and it indicates that the NALT is the site of antigen-specific T-cell priming.

scholarly journals Identification of LAG3+ T Cell Populations in the Tumor Microenvironment of Classical Hodgkin Lymphoma and B-Cell Non-Hodgkin Lymphoma

Blood ◽  
2020 ◽  
Vol 136 (Supplement 1) ◽  
pp. 19-19
Author(s):  
Katsuyoshi Takata ◽  
Tomohiro Aoki ◽  
Lauren C. Chong ◽  
Katy Milne ◽  
Tomoko Miyata-Takata ◽  
...  
Keyword(s):  
T Cells ◽  
T Cell ◽  
B Cell ◽  
Hodgkin Lymphoma ◽  
Research Funding ◽  
Cell Lymphoma ◽  
Expression Patterns ◽  
B Cell Lymphoma ◽  
Cell Populations ◽  
Classical Hodgkin Lymphoma

Background: LAG3 is one of the immune check point receptors that are expressed on activated cytotoxic T-cells and regulatory T cells. Physiologically, T-cell proliferation and memory T-cell differentiation is negatively regulated by LAG3-MHC interaction. In cancer tissues, T-cells that are chronically exposed to tumor antigens might upregulate LAG3 and receive inhibitory stimuli to enter an exhaustion state limiting anti-tumor immune responses. Currently, clinical trials using double blockade of LAG3/PD1 are active in several solid tumours, but there are only a small number of clinical trials using LAG3 monoclonal antibodies in lymphoma. Recently, we published a characteristic LAG3+ T-cell population as a mediator of immune suppression in classical Hodgkin lymphoma (Aoki & Chong et al. Cancer Discovery 2020). However, the abundance and variability of LAG3 positive T-cell populations across a spectrum of B-cell lymphoma has not been well studied and it remains an open question if LAG3 expression is associated with treatment outcome under standard-of-care conditions. Methods: We performed a LAG3 immunohistochemical (IHC) screen in a large cohort of B-cell Non-Hodgkin lymphoma (diffuse large B-cell lymphoma (DLBCL); N=341, follicular lymphoma (FL); N=198 (grade 1-3A), transformed FL to aggressive lymphoma (tFL); N=120, mantle cell lymphoma (MCL); N=179, primary mediastinal large B-cell lymphoma (PMBCL); N=61) and classical Hodgkin lymphoma (HL; N=459) to assess LAG3 expression in the tumor microenvironment (TME). Moreover, we characterized LAG3+ T-cell populations using multi-color immmunohistochemistry (IHC) (LAG3, PD1, CD4, CD8, FOXP3, CD20) in various lymphoma subtypes. Clinical parameters including treatment outcome were correlated with the abundance of LAG3+ T-cell populations in the TME. Results: On average, HL (7%) and PMBCL (6%) showed higher LAG3+ cellular frequency than the other B-cell lymphoma subtypes studied (DLBCL and FL: 2%, MCL: 0.8%). Comparing the frequency of LAG3+ cells according to MHC class I/II status, DLBCL showed a significant correlation with MHC class I status, and LAG3 expression correlated with MHC class II status in HL. Next, we performed multi-color IHC to describe subtype-specific expression patterns of LAG3 in T cell subsets. LAG3+PD1- T-cells were predominantly found in HL and PMBCL with only rare LAG3+PD1+ cells in HL. The majority of LAG3+ T-cells co-expressed CD4 in HL, in contrast to CD8 in PMBCL. DLBCL showed a mixed population pattern with a 1:1 ratio of LAG3+PD1- and LAG3+PD1+ T-cells. In FL, the majority of LAG3+ T-cells were CD4+PD1+, suggesting a more exhausted TME phenotype in FL than in other lymphoma subtypes. Cellular distance analysis showed that LAG3+CD4+ T-cells were in close vicinity to CD20+ lymphoma cells in FL, while in DLBCL and PMBCL, the nearest neighbors of malignant cells were LAG3+CD8+. Triple-positive LAG3+PD1+CD8+ T-cells significantly correlated with high infiltrating M2 macrophage (Pearson's correlation test, P &lt; 0.001) content and the ABC cell-of-origin subtype (Pearson's correlation test, P = 0.002) in DLBCL. The abundance of LAG3+CD8+PD1- cells correlated with a high FLIPI score (Pearson's correlation test, P = 0.033), disease specific survival (HR = 2.8, 95% CI = 1.3-5.9, P = 0.006), time to progression (HR = 2.8, 95% CI = 1.6-5.0, P = 0.001) and transformation (HR = 4.0, 95%CI = 1.7-9.6, P = 0.002) in FL treated with R-CVP (N = 135). Assessing LAG3 expression by single color IHC in FL (cut-off at 5%), patients with LAG3-positive samples showed significantly higher FL transformation rates (P = 0.023) and tFL samples showed higher abundance of LAG3+ cells than the corresponding primary pretreatment FL samples (primary FL: 1.5±1.7% vs. tFL: 4.2±3.8%, t-test, P = 0.01). The increased transformation risk was validated in an independent FL cohort treated with R-CHOP/CVP (N=97, HR = 6.2, 95% CI = 2.8-13.9, P &lt; 0.001). Conclusion: The highest abundance of LAG3+ T-cells in the TME was found in HL and its related entity PMBCL. The differential outcome correlations and co-expression patterns in LAG3+ T cells across B-cell lymphoma subtypes indicate heterogeneity in TME composition and related pathogenic mechanisms. Our results suggest that LAG3 expression patterns will be important in the interpretation of ongoing studies and highlight populations that may benefit from LAG3 checkpoint inhibition. Disclosures Sehn: AstraZeneca: Consultancy, Honoraria; Genentech, Inc.: Consultancy, Honoraria, Research Funding; Amgen: Consultancy, Honoraria; AbbVie: Consultancy, Honoraria; Chugai: Consultancy, Honoraria; TG therapeutics: Consultancy, Honoraria; Verastem Oncology: Consultancy, Honoraria; Teva: Consultancy, Honoraria, Research Funding; Servier: Consultancy, Honoraria; F. Hoffmann-La Roche Ltd: Consultancy, Honoraria, Research Funding; MorphoSys: Consultancy, Honoraria; Takeda: Consultancy, Honoraria; Apobiologix: Consultancy, Honoraria; Seattle Genetics: Consultancy, Honoraria; Gilead: Consultancy, Honoraria; Kite: Consultancy, Honoraria; Merck: Consultancy, Honoraria; Lundbeck: Consultancy, Honoraria; Karyopharm: Consultancy, Honoraria; Janssen: Consultancy, Honoraria; Celgene: Consultancy, Honoraria; Acerta: Consultancy, Honoraria. Savage:Merck, BMS, Seattle Genetics, Gilead, AstraZeneca, AbbVie, Servier: Consultancy; BeiGene: Other: Steering Committee; Roche (institutional): Research Funding; Merck, BMS, Seattle Genetics, Gilead, AstraZeneca, AbbVie: Honoraria. Scott:Celgene: Consultancy; Abbvie: Consultancy; AstraZeneca: Consultancy; NIH: Consultancy, Other: Co-inventor on a patent related to the MCL35 assay filed at the National Institutes of Health, United States of America.; Roche/Genentech: Research Funding; NanoString: Patents & Royalties: Named inventor on a patent licensed to NanoString, Research Funding; Janssen: Consultancy, Research Funding. Steidl:Bayer: Consultancy; Juno Therapeutics: Consultancy; Roche: Consultancy; Seattle Genetics: Consultancy; Bristol-Myers Squibb: Research Funding; AbbVie: Consultancy; Curis Inc: Consultancy.

scholarly journals Quantitative Analysis of the Acute and Long-Term CD4+ T-Cell Response to a Persistent Gammaherpesvirus

1999 ◽  
Vol 73 (5) ◽  
pp. 4279-4283 ◽  
Author(s):  
Jan P. Christensen ◽  
Peter C. Doherty
Keyword(s):  
T Cells ◽  
T Cell ◽  
Lymph Nodes ◽  
Cell Response ◽  
Peak Frequency ◽  
T Cell Response ◽  
Compensatory Response ◽  
Cervical Lymph Nodes ◽  
Murine Gammaherpesvirus

ABSTRACT The murine gammaherpesvirus 68 (MHV-68) replicates in respiratory epithelial cells, where it establishes a persistent, latent infection limited predominantly to B lymphocytes. The virus-specific CD4+ T-cell response in C57BL/6 mice challenged intranasally with MHV-68 is detected first in the mediastinal lymph nodes and then in the cervical lymph nodes and the spleen. The numbers of MHV-68-specific CD4+ T cells generated in congenic mice homozygous for disruption of the β2-microglobulin gene tended to be higher, indicating that the absence of the CD8+ set in this group resulted in a compensatory response. The peak frequency within the splenic CD4+ T-cell population may reach 1:50 in the acute response; it then drops to 1:400 to 1:500 within 4 months and stays at that level in the very long term. Sorting for L-selectin (CD62L) expression established that all virus-specific CD4+ T cells were initially CD62Llow, with >80% maintaining that phenotype for the next 14 months. The overall conclusion is that MHV-68-specific CD4+ T cells remain activated (CD62Llow) and at a stable frequency in the face of persistent infection.

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