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1.  Combination chemotherapy with carboplatin, capecitabine and epirubicin (ECarboX) as second- or third-line treatment in patients with relapsed ovarian cancer: a phase I/II trial 
British Journal of Cancer  2006;94(1):74-78.
Platinum-based combination chemotherapy has been proven to be superior to single-agent platinum in the treatment of relapsed ovarian cancer after a treatment-free interval of more than 6 months. A response rate of 41% was previously reported by our group using a combination of epirubicin, cisplatin and 5-FU in patients who relapsed within 12 months, we therefore assessed a similar, but more convenient combination of epirubicin, carboplatin and capecitabine in this phase-I/II trial. In total, 18 patients with recurrent epithelial ovarian carcinoma, who had not received more than two lines of chemotherapy and the treatment-free interval exceeded 6 months were treated with carboplatin AUC5, epirubicin 50 mg m−2 and capecitabine at several dose levels on continuous 21 day cycles and 14 of 21 day cycles. Patients were assessed for toxicity and by CT and CA-125 for response. The overall response rate was 61.1%, with three complete and eight partial responses. Grade 3/4 haematological toxicity was seen in 10 out of 18 patients and caused dose reductions and treatment delays. The combination of epirubicin, carboplatin and capecitabine showed good activity but caused excessive toxicity. A phase-II trial using carboplatin and capecitabine is underway.
doi:10.1038/sj.bjc.6602879
PMCID: PMC2361084  PMID: 16306873
capecitabine; carboplatin; epirubicin; ovarian cancer; relapse
2.  Cytotoxic T-Lymphocyte Responses to a Polymorphic Epstein-Barr Virus Epitope Identify Healthy Carriers with Coresident Viral Strains 
Journal of Virology  2000;74(4):1801-1809.
Cytotoxic T-lymphocyte (CTL) responses to Epstein-Barr virus (EBV) tend to focus on a few immunodominant viral epitopes; where these epitope sequences are polymorphic between EBV strains, host CTL specificities should reflect the identity of the resident strain. In studying responses in HLA-B27-positive virus carriers, we identified 2 of 15 individuals who had strong CTL memory to the pan-B27 epitope RRIYDLIEL (RRIY) from nuclear antigen EBNA3C but whose endogenous EBV strain, isolated in vitro, encoded a variant sequence RKIYDLIEL (RKIY) which did not form stable complexes with B27 molecules and which was poorly recognized by RRIY-specific CTLs. To check if such individuals were also carrying an epitope-positive strain (either related to or distinct from the in vitro isolate), we screened DNA from freshly isolated peripheral blood mononuclear cells for amplifiable virus sequences across the EBNA3C epitope, across a different region of EBNA3C with type 1-type 2 sequence divergence, and across a polymorphic region of EBNA1. This showed that one of the unexplained RRIY responders carried two distinct type 1 strains, one with an RKIY and one with an RRIY epitope sequence. The other responder carried an RKIY-positive type 1 strain and a type 2 virus whose epitope sequence of RRIFDLIEL was antigenically cross-reactive with RRIY. Of 15 EBV-seropositive donors analyzed by such assays, 12 appeared to be carrying a single virus strain, one was coinfected with distinct type 1 strains, and two were carrying both type 1 and type 2 viruses. This implies that a small but significant percentage of healthy virus carriers harbor multiple, perhaps sequentially acquired, EBV strains.
PMCID: PMC111658  PMID: 10644353
3.  Epstein-Barr Virus Nuclear Antigen 1 Sequences in Endemic and Sporadic Burkitt’s Lymphoma Reflect Virus Strains Prevalent in Different Geographic Areas 
Journal of Virology  1999;73(2):965-975.
The Epstein-Barr virus (EBV) nuclear antigen EBNA1 is the only viral protein detectably expressed in virus genome-positive Burkitt’s lymphoma (BL); recent work has suggested that viral strains with particular EBNA1 sequence changes are preferentially associated with this tumor and that, within a patient, the tumor-associated variant may have arisen de novo as a rare mutant of the dominant preexisting EBV strain (K. Bhatia, A. Raj, M. J. Gutierrez, J. G. Judde, G. Spangler, H. Venkatesh, and I. T. Magrath, Oncogene 13:177–181, 1996). In the present work we first study 12 BL patients and show that the virus strain in the tumor is identical in EBNA1 sequence and that it is matched at several other polymorphic loci to the dominant strain rescued in vitro from the patient’s normal circulating B cells. We then analyze BL-associated virus strains from three different geographic areas (East Africa, Europe, and New Guinea) alongside virus isolates from geographically matched control donors by using sequence changes in two separate regions of the EBNA1 gene (N-terminal codons 1 to 60 and C-terminal codons 460 to 510) to identify the EBNA1 subtype of each virus. Different geographic areas displayed different spectra of EBNA1 subtypes, with only limited overlap between them; even type 2 virus strains, which tended to be more homogeneous than their type 1 counterparts, showed geographic differences at the EBNA1 locus. Most importantly, within any one area the EBNA1 subtypes associated with BL were also found to be prevalent in the general population. We therefore find no evidence that Burkitt lymphomagenesis involves a selection for EBV strains with particular EBNA1 sequence changes.
PMCID: PMC103916  PMID: 9882297
4.  Bolus/infusional 5-fluorouracil and folinic acid. A report on two prospective, consecutive phase II studies with 5-fluorouracil dose escalation. 
British Journal of Cancer  1998;77(9):1480-1486.
We have used a relatively new trial methodology, the group sequential design, to prospectively evaluate two dose levels of bolus/infusional 5-fluorouracil (5-FU) and folinic acid in 192 consecutive-patients with advanced colorectal carcinoma. On day 1, all patients received 200 mg m(-2) of folinic acid infusion over 2 h. Cohort A (n = 102 patients) received 500 mg m(-2) 5-FU by i.v. 15-min infusion followed by an infusion of 500 mg m(-2) 5-FU over 22 h. Treatment was repeated on day 2 and further cycles given 2-weekly. After sequential analysis excluded a response rate of over 40%, cohort B (n = 90 patients) received an increased dose of 600 mg m(-2) 5-FU bolus and infusion. Patients had received no prior 5-FU therapy and the two cohorts had similar demographic features. In 179 evaluable patients, the overall response rate was 18% (95% CI 12-24%) with CR of 6% and PR of 12%, with no difference between the two cohorts. Overall median survival was 34 weeks (95% CI 30-39) with no significant difference between cohorts (median survival 32 and 37 weeks in cohort A and B respectively; P = 0.27). On multivariate analysis, poor performance status, elevated initial WBC and alkaline phosphatase and low serum albumin were associated with reduced survival (P < 0.05), and initial raised WBC showed an association with reduced likelihood of response (P = 0.002). Overall toxicity was low with CTC grade 3 mucositis, diarrhoea, nausea or vomiting in < or = 6% of patients and no treatment-related deaths. Significant (grade 3 or above) leucopenia was more common in cohort B than in cohort A (9% and 1% respectively); there were more dose reductions, and the median administered dose intensity was lower in cohort B than in cohort A (89% and 97% respectively; P = 0.006). In this group of relatively unselected patients, we have confirmed a relatively low objective response rate and median survival of 7.8 months with this regimen. There was no significant difference in outcome between the two dose levels but the higher dose of 5-FU was associated with more dose reductions and greater toxicity.
PMCID: PMC2150187  PMID: 9652765
5.  Epidemiology of Infection with Epstein-Barr Virus Types 1 and 2: Lessons from the Study of a T-Cell-Immunocompromised Hemophilic Cohort 
Journal of Virology  1998;72(5):4352-4363.
In apparent contrast to earlier work on Epstein-Barr virus (EBV) carriage in the general Caucasian population, in vitro virus isolations from human immunodeficiency virus (HIV)-positive male homosexual cohorts have shown frequent examples of multiple EBV infection and an overall prevalence of type 2 EBV strains exceeding 30%. Here we ask to what extent these findings might hold true in another T-cell-immunocompromised cohort, HIV-positive hemophilic patients. Resident EBV strains were rescued within lymphoblastoid cell lines derived from the blood and throat washings of 39 such individuals, using the same in vitro protocols of virus isolation as for the homosexual cohort. A mean of 19 independent cell lines was made per patient, and in each case the resident virus was characterized by PCR-based viral genomic analysis and by immunoblotting to reveal the viral “EBNAprint.” By these criteria a significant proportion (14 of 39) of the hemophilic cohort carried more than one EBV strain, suggesting that T-cell impairment does indeed sensitize virus carriers to reinfection with new strains of exogenously transmitted virus. However, the overall incidence of type 2 EBV infection was 10%, which is close to that observed in the earlier work with healthy carriers and substantially lower than that seen in HIV-positive homosexuals. We infer that type 2 EBV is relatively rare in the general Caucasian population but has become endemic in the homosexual community.
PMCID: PMC109665  PMID: 9557725
6.  Phase I study of accelerated FEC with granulocyte colony-stimulating factor (Lenograstim) support. 
British Journal of Cancer  1995;71(6):1279-1282.
With the aim of increasing the dose intensity of chemotherapy in breast cancer, 14 patients with stage II-IV breast cancer were treated with FEC chemotherapy at 2 week intervals together with granulocyte colony-stimulating factor (G-CSF) 5 micrograms kg-1 s.c. on days 2-14. Five of six patients completed six courses of 5-fluorouracil 600 mg m-2, epirubicin 60 mg m-2 and cylcophosphamide 600 mg m-2 within 11 weeks. Eight patients were treated with 5-fluorouracil 700 mg m-2, epirubicin 70 mg m-2 and cyclophosphamide 700 mg m-2 and four had dose-limiting toxicity with sepsis, thrombocytopenia or mucositis. All patients who received G-CSF had satisfactory neutrophil counts by day 15 of each course. Cumulative anaemia and thrombocytopenia were observed, but treatment at the first dose was tolerable. Seven of eight patients with measurable disease had partial responses. This regimen permits a 50% increase in dose intensity compared with conventional treatment at 3 week intervals and warrants further evaluation.
PMCID: PMC2033830  PMID: 7540038
7.  The prognostic value of a response to chemotherapy given before radiotherapy in advanced cancer of cervix. 
British Journal of Cancer  1989;59(3):473-475.
Forty patients with stage III and 15 patients with stage IVa carcinoma of cervix have been treated with two pulses of cisplatin, vincristine and bleomycin combination chemotherapy before full dose radical radiotherapy. Twenty-seven of 51 (53%, 95% confidence interval 40-67%) had an objective response to chemotherapy and all chemotherapy responders had a complete response to radiotherapy. The actuarial survival at 24 months of responders to chemotherapy is 71% against 37% for non-responders. The responding patients had an estimated reduction in mortality to 36% (P = 0.014, 95% CI 15-81%) of that of the non-responders to chemotherapy. The incidence of tumour recurrence or distant metastases showed a similar reduction to 34% (P = 0.006, 95% CI 14-73%) of that of the non-responders. The data strongly suggest that response to chemotherapy in the initial treatment of advanced cervical cancer is associated with an improvement in survival following subsequent radical radiotherapy.
PMCID: PMC2247076  PMID: 2467687
8.  Angio-immunoblastic lymphadenopathy: a clinical, immunological and molecular study. 
British Journal of Cancer  1987;55(4):437-442.
Twenty four patients with angio-immunoblastic lymphadenopathy (AILD) presenting between 1974 and 1985 have been reviewed. Clinical features at presentation included rash, fever, lymphadenopathy and hepatosplenomegaly in 75% of patients. Polyclonal hypergammaglobulinaemia was seen in 19/20 patients; 5 had normal immunoglobulin levels. Combination chemotherapy with MVPP was the optimal treatment with 6/7 patients achieving complete remission. Duration of remission ranged from 9 months to 4 years and was significantly longer in patients achieving complete as opposed to partial remission. In 6 patients phenotype studies were performed on single cell suspensions and immunoperoxidase studies on frozen sections of 7 lymph nodes. There was a reversal of the normal T suppressor/helper cell ratio with a predominance of T suppressor cells. Loss of normal B follicles was observed histologically in all except one lymph node. Germline configuration of the beta B-chain of the T cell receptor was observed in lymph nodes of 4 patients with AILD, and a rearranged T cell receptor was observed in 1 patient in whom a second lymph node biopsy had shown alteration of the histological picture to that of T-zone lymphoma. Frozen sera of 15 patients were screened for antibodies to HTLV I and III and were found to be negative.
PMCID: PMC2001687  PMID: 2953385
9.  New marker of B lymphocytes, MB2: comparison with other lymphocyte subset markers active in conventionally processed tissue sections. 
Journal of Clinical Pathology  1987;40(2):151-156.
The use of the murine monoclonal antibody MB2 for identifying B lymphocytes in routinely processed tissue was evaluated and contrasted with the use of the monoclonal antibody UCHL1 for identifying T cells. One hundred and sixty eight surgical biopsy specimens were immunostained with these antibodies, including a wide range of normal and neoplastic non-lymphoid tissues, as well as normal lymphoreticular tissues and lymphomas. Sixty four non-Hodgkin's lymphomas were also examined, of which 51 had been previously phenotypically defined. In selected cases the results were compared with those obtained using two other monoclonal antibodies MB1 and MT1, used for identifying B and T cells, respectively, in paraffin sections. MB1 stained a smaller proportion of B cell tumours than MB2 and staining was, in general, weaker, except in one case of centroblastic lymphoma. MT1 immunoreactivity was comparable with that of UCHL1, except in one case of T lymphoblastic lymphoma (MT1 positive, UCHL1 negative). None of the antibodies is ideal, but, if used as a panel, they permit the separation of B cells and T cells in paraffin sections.
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PMCID: PMC1140858  PMID: 3546393
10.  Normal values for the different classes of venous blood mononuclear cells defined by monoclonal antibodies. 
Journal of Clinical Pathology  1985;38(1):54-59.
We present the data obtained from routine quantitation of normal venous blood mononuclear cells using 19 monoclonal antibodies against defined mononuclear cell surface antigens. The results indicate different values for percentage and absolute numbers of T lymphocytes and of T lymphocyte subsets depending on the monoclonal antibodies used to quantify these populations. In most instances the total number of identified cells was significantly less than the total number of recovered blood mononuclear cells, which suggests the existence in blood of a null cell population. Evidence is advanced to support previous observations that at least a proportion of this population is of B cell lineage, expressing cytoplasmic immunoglobulin but lacking other class or lineage specific markers. The value of a diverse monoclonal panel in routine quantitation of peripheral blood mononuclear cells is discussed.
PMCID: PMC499071  PMID: 3871442
11.  Lymphocytic lymphoma/B-chronic lymphocytic leukaemia--an immunohistopathological study of peripheral B lymphocyte neoplasia. 
British Journal of Cancer  1984;50(5):587-599.
Twenty seven patients with malignant lymphoma of lymphocytic type/B-chronic lymphocytic leukaemia (ML, L/B-CLL, Kiel classification) diagnosed from lymph node and splenectomy specimens were studied histologically and immunologically. Lymph node biopsies showed a diffuse effacement of normal architecture by small round lymphocytes usually with scattered proliferation centres (PC). A1 spleens showed white pulp with or without red pulp involvement, sometimes with tumour nodules present. PC-like cells or PC were only found in the white pulp or tumour nodules. Studies of 13 specimens using the ABC immunoperoxidase technique on frozen sections with a large panel of monoclonal antibodies showed that although a part of the monoclonal B cell neoplasm, the proliferation centres or splenic white pulp have a different phenotype from the surrounding cells. Some of these phenotypic changes are similar to those reported with in vitro induction of "maturation" of ML, L/B-CLL cells. The implications for normal B-cell development are discussed. In contrast to reported peripheral blood findings, T cells, predominantly of T helper phenotype in lymph nodes, were present but usually not numerous.
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PMCID: PMC1976971  PMID: 6388614
12.  Characteristics of Sternberg-Reed, and related cells in Hodgkin's disease: an immunohistological study. 
British Journal of Cancer  1984;49(4):465-476.
A panel of monoclonal antileucocyte antibodies was used in a study of Hodgkin's disease (HD) to explore the phenotypic characteristics of Sternberg-Reed and related cells (collectively termed HD cells). Cryostat preparations of 31 lymph nodes and 2 spleens were obtained from 30 patients with active HD. The histological diagnoses were: lymphocyte predominance (LP), 4 patients; nodular sclerosis (NS), 22; mixed cellularity (MC), 2; lymphocyte depletion (LD), 2. The monoclonal antibodies used were: OKT3, T11, Leu-1 (pan T cell specific); Leu-3A (T "helper" specific); Leu-2A, OKT8 (T "suppressor" specific); immunoglobulin (Ig) antibodies: anti kappa and lambda light chains, anti mu and delta heavy chains; B1 (anti B lymphocyte); CA2-11 (anti HLA-DR); OKM1, Mo-2 (anti myeloid/monocyte); OKT9 (anti transferrin receptor); Leu-7 (anti "NK" cell) and J5 (anti common ALL antigen). Reactions with peanut lectin (PNL) were also studied. The reactions were developed using a modified "ABC" immunoperoxidase technique. Specific attention was paid to the cell surface phenotype and anatomical localisation of HD cells in relation to surrounding T and B lymphocytes. HD cells formed distinct "rosettes" with T cells of "helper" phenotype although in 3 cases (1: LP, 2: NS) Leu-7 positive cells formed a prominent component of these interactions. In partially involved lymph node and spleen, HD cells were prominently distributed in a perifollicular distribution. In addition follicular mantle zones were frequently infiltrated by HD cells, the degree of ensuing destruction being related to the extent of lymph node effacement by HD. In 2 cases (1: NS, 1: LD) HD cells expressed clear, positive reactions with B1 although in neither of these cases nor in any other instance, was surface Ig expressed on the HD cell surface. The great majority of HD cells reacted positively with both OKT9 and, as previously reported, with anti HLA-DR antibody. In addition, HD cells demonstrated intense surface and cytoplasmic staining with PNL. HD cells were negative with all other antibodies. On the basis of these findings, no lineage specificity can confidently be attributed to the HD cell. However, the pattern of immunohistological reactions suggest that it is related to a cell of B follicular origins.
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PMCID: PMC1976777  PMID: 6370286
13.  Early involvement of the bronchi in patients with malignant lymphoma. 
British Journal of Cancer  1983;48(6):777-781.
Fibreoptic bronchoscopy in previously untreated patients with malignant lymphoma provided diagnostic information in 8 of 25 cases with radiological evidence of intrathoracic involvement. There was a marked difference in the pattern of endobronchial involvement between Hodgkin's Disease (HD) and Non Hodgkin's Lymphoma (NHL) which specifically infiltrated the bronchus-associated lymphoid tissue. Bronchoalveolar lavage was abnormal in only one patient with Hodgkin's Disease in whom the presence of many Sternberg-Reed cells suggested occult dissemination of otherwise localised disease.
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PMCID: PMC2011569  PMID: 6652018
14.  Paradoxical response of malignant melanoma to methotrexate in vivo and in vitro. 
British Journal of Cancer  1983;47(5):671-679.
Methotrexate (MTX) shows consistent cytotoxicity for melanoma cells in vitro but it is ineffective in clinical use at equivalent concentrations in vivo. This apparent paradox has been investigated by cell culture techniques and results quantified by cell number. In an in vitro model of high dose MTX therapy followed by leucovorin rescue (HD-MTX-LCR) there was survival of both melanoma and choriocarcinoma cell lines but not of an acute lymphocytic leukaemia cell line. The 70H metabolite of MTX was identified by HPLC in plasma samples of melanoma patients treated by HD-MTX-LCR, in which MTX concentrations approximately 10(-5) M were maintained for 24 h. However, metabolism per se is unlikely to account for the lack of response to MTX clinically. In vitro 70H MTX (10(-7) - 10(-6) M) was two orders of magnitude less cytotoxic for melanoma than MTX (10(-9) - 10(-8) M). The cellular accumulation of [3H]-MTX, using a rapid gradient centrifuge technique for separation of melanoma cells from medium, was reduced in the presence of 70H-MTX. The results suggest that reduced cellular uptake of MTX combined with biochemical rescue of tumour cells may partially explain the paradoxical lack of clinical response of melanoma to the drug.
PMCID: PMC2011395  PMID: 6601959
15.  The cellular content of non Hodgkin lymphomas: a comprehensive analysis using monoclonal antibodies and other surface marker techniques. 
British Journal of Cancer  1983;47(3):327-351.
Five samples of tonsil, 10 reactive lymph nodes and 65 consecutive cases of non-Hodgkin lymphoma (NHL) were evaluated in suspension phenotyping with the monoclonal antibodies alpha Leu-I, alpha Leu-2a, alpha Leu-3a, OKT1, OKT3, OKT4, OKT6, OKT8, W6/32, 26/114, DA-2, 2DI, J5, AN51 and OKT9 together with conventional surface marking by rosetting (E, Fc gamma, Fc mu, C3b, C3d) and staining for surface and cytoplasmic immunoglobulin (SIg, CyIg) heavy and light chain classes. The results confirm the reproductability and specificity of staining with monoclonal antibodies against T cells and T cells subsets. Evidence is presented for reactivity of alpha Leu-I antibody with SIg positive and Ia positive cells in some lymphomas (centroblastic centrocytic, lymphocytic and immunoblastic), and 2 cases showed evidence of marking with OKT3 on SIg positive cells in T cell predominant immunoblastic lymphoma. Lymphoblastic lymphomas of T cell type expressed the marker OKT6. On the basis of these results criteria for the diagnosis of T cell lymphoma are suggested. The monoclonal antibody J5, reactive with C-ALL antigen, showed variable positivity, occasionally strong in B cells in cases of centroblastic and centrocytic follicular lymphoma. Proportions of cells staining with the monoclonal antibody OKT9 showed a correlation between levels of cellular expression of transferrin (trf) receptor and the histological grade of malignancy, OKT9+ cells being elevated in high grade lymphomas, and in some cases of transforming lymphoma of low grade histological class. These results are discussed and indicate the advantage of employing a wide range of defined monoclonal reagents in the phenotypic evaluation of NHL.
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PMCID: PMC2011301  PMID: 6338896
16.  Distribution of T-lymphocyte subsets in Hodgkin's disease characterized by monoclonal antibodies. 
British Journal of Cancer  1982;45(4):491-499.
Mononuclear-cell suspensions of lymph nodes, spleen and blood from 24 patients with active Hodgkin's disease (HD) were studied for possible imbalance of T and B lymphocytes, and T-lymphocyte subsets, using monospecific anti-T antibodies and other reagents. A profile showing T-cell predominance was demonstrated in lymph nodes and blood, with total T-cells ranging from 50-70% of the cell count. As defined by monoclonal antibodies, 70-85 of the latter comprised the "inducer" subclass, the remainder being "suppressor" cells. There were no essential differences between histologically involved and uninvolved lymph nodes from HD patients, though total T-cell proportions were lower in "normal lymph node" controls. The profiles of spleens electively removed, as part of pre-treatment staging procedures, showed reduced total T-cell numbers, whether these were involved with HD or not. These differences are accounted for principally by fewer T "inducer" cells (24%, in spleen, v. 54% in involved lymph nodes and 47% in "normal" control nodes). Possible explanations for these findings are discussed. Our results demonstrate similar profiles in histologically diseased and normal tissue, rather than any clear imbalance of T-cell proportions which might explain the profound disturbances of T-cell function frequently demonstrated in vivo and in vitro.
PMCID: PMC2010995  PMID: 6978728
17.  Binding of peanut lectin to germinal-centre cells: a marker for B-cell subsets of follicular lymphoma? 
British Journal of Cancer  1981;44(1):68-74.
The binding of horseradish-peroxidase-labelled peanut lectin (HRP-PNL) to cryostat sections of tonsil, lymphoma lymph nodes, reactive lymph nodes and miscellaneous tumours demonstrated that PNL binds selectively to lymphocytes in germinal centres. Lymph nodes from 21 patients with non-Hodgkin's lymphomas were phenotyped as cell suspensions for PNL binding, and the following surface markers: E rosetting, C3d, SIg, OK markers of T-cell subsets, Ig heavy-chain and light-chain classes. There was a positive correlation between PNL binding and cells with SIg and C3d receptors. 4/5 cases of centroblastic/centrocytic follicular lymphoma had a PNL+ SIg+ C3d+ phenotype. Both cases of centroblastic/centrocytic diffuse lymphoma were PNL-. There was no correlation between PNL binding and heavy- or light-chain Ig class. PNL binding and presence of C3d receptors were not always positively correlated, indicating that follicular cells may be either PNL+ SIg+ C3d+ or PNL+ SIg+ C3d-. The binding pattern of PNL to 1 case of thymic hyperplasia and 2 cases of malignant lymphoma lymphoblastic T type suggested that some but not all cortical thymocytes bind PNL.
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PMCID: PMC2010668  PMID: 6789856
18.  Immuno-histological diagnosis of lymphoproliferative diseases by selected combinations of antisera and monoclonal antibodies. 
British Journal of Cancer  1980;42(2):224-242.
Tissue sections of frozen biopsy specimens obtained from normal and hyperplastic human lymphoid tissues, 33 cases of non-Hodgkin lymphomas as well as various forms of immunoregulatory disorders (angioimmunoblastic and dermatopathic lymphadenopathy) were analysed in immunofluorescence tests (using red TRITC and green FITC double-labelling). A panel of antisera including well-characterized conventional reagents to immunoglobulin classes, T lymphoid and Ia-like antigens, and monoclonal antibodies was used. In selected cases the results were compared with the observations of membrane-marker staining on viable cells in suspension. the findings show that the immunological methods can give a very accurate analysis of the normal and malignant lymphoid cells, and can provide complementary information to conventional histology. The investigator can choose the reagent combinations which give answers to various specific questions: e.g. antisera to light chains establish the monoclonality of lymphomas, whilst staining combinations for human T and Ia-like antigens are particularly useful in various immunoregulatory disorders. Monoclonal antibodies will be particularly useful in various immunoregulatory disorders. Monoclonal antibodies will be particularly useful reagents for analysing the tissue distribution of lymphoid subpopulations and ancillary cells in tissue biopsy specimens.
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PMCID: PMC2010398  PMID: 6775656
19.  Surface phenotyping, histology and the nature of non-Hodgkin lymphoma in 157 patients. 
British Journal of Cancer  1979;40(1):11-34.
In a study of 157 patients with lymphoid malignancy, the phenotype of the tumour cells was correlated with the histological classification of the tumour using the Rappaport and the Kiel classifications. The markers used included E, Fc gamma, Fc micron (IgM) and C3d rosetting, estimation of SIg and CyIg, and tests for the expression of HTLA, Ia and ALL. Repeat biopsy specimens were studied in 23 of these patients. The phenotypic features of lymphoblastic malignancy indicated B-cell, T-cell and ALL-positive null-cell tumours in this group. Immunoblastic lymphomas were predominantly of non-capping B-cell type, but T-cell immunoblastic lymphoma occurred in 2 patients. Immunoblastic lymphomas of receptor-silent cells occur, and are ALL- and HTLA-negative. In the category of diffuse, poorly differentiated lymphocytic lymphomas, most cases are of centroblastic and centrocytic tumour of diffuse type, but pure centrocytic tumours and centroblastic tumours occur. The dominant phenotype in this group is of B cells expressing C3d receptors. Nodular poorly differentiated lymphocytic lymphomas (Rappaport) are classified as centroblastic and centrocytic follicular (Kiel) and most express SIg+ C3d+ phenotype. The frequency of this phenotype appeared the same in both diffuse and nodular poorly differentiated lymphocytic neoplasms. The Rappaport group of diffuse well-differentiated lymphocytic lymphoma includes 2 Kiel categories, malignant lymphoma lymphocytic, and malignant lymphoma lymphoplasmacytoid. Cells of the former tumour were considered to be immature B cells resembling those seen in CLL, and characteristically expressing SIg weakly, with a high frequency of single kappa light chain. Cells of the latter tumour are by contrast mature, and are related to the centroblastic and centrocytic follicular tumour by their histogenesis and phenotypic features. Repeat biopsy examinations indicate that T-cell predominance occurs in the prodromal phase of B-cell-predominant tumours of SIg+ C3d+ phenotype. It is concluded that non-Hodgkin lymphoma can be divided into 2 categories: (1) tumours of immature immunologically incompetent cells of lymphoblastic histology and with phenotypic features akin to T, B and Null-cell ALL, and (2) tumours of differentiated lymphocytes expressing the phenotypic features of B lymphocytes, with maturation arrested at one of several stages of an antigen-dependent immune response.
PMCID: PMC2009956  PMID: 314300
20.  Terminal deoxynucleotidyl transferase activity in lymphoma. 
British Journal of Cancer  1979;39(5):566-569.
Terminal deoxynucleotidyl transferase (TdT) was estimated in the tissues of 42 patients with lymphoma, whose cells were also typed by the use of surface markers. Four of the 8 patients with T-cell lymphoma were TdT+ including patients whose lymph nodes showed an undifferentiated or poorly differentiated appearance. The TdT- T-cell lymphomas included cases with diffuse histiocytic Sezary cell, diffuse, poorly differentiated and angio-immunoblastic histology. The tissues of 31 patients with B-cell lymphoma were invariably TdT-, whether the histology was poorly differentiated, well differentiated, nodular, diffuse, histiocytic or Burkitt type, and including cases with about equal proportions of T and B cells, and those whose cells showed non-capping and capping surface immunoglobulin. Hodgkin's tissue was also invariably TdT-. We conclude that estimation of TdT in tissues of patients with malignant lymphoma may be a useful test in diagnosing the T-cell lymphoma, particularly in patients with tumours of undifferentiated or poorly differentiated histology.
PMCID: PMC2009895  PMID: 314814
21.  Correlation of surface receptors with histological appearance in 29 cases of non-Hodgkin lymphoma. 
British Journal of Cancer  1977;35(6):858-867.
The receptor patterns of cell suspensions from 29 cases of non-Hodgkin lymphoma were correlated with the histology of the nodes from which the cells were taken. Twenty-two were judged to be predominantly or largely B-cell, and because of this preponderance these were divided by a method based on the distribution of surface immunoglobulin and the expression of Fc and C3 receptors. "Mature" B-cell and B-mixed tumours showing capping surface Ig with Fc and/or C2 receptors correlated well with a nodular growth pattern, and consisted of what Rappaport (1966) calls "poorly differentiated" lymphocytes equivalent to the "small cleaved" cells as defined by Lukes and Collins (1975). Ten of the 14 patients in this receptor category are alive between 12 and 30 months after diagnosis. Receptor-silent and "immature" B-cell tumours with non-capping surface Ig correlated predominantly with the Rappaport histiocytic lymphoma and Lukes and Collins' large cleaved and large non-cleaved lymphomas, though these histological categories also included a wide variety of other receptor types such as T-cell, Receptor-overlap and the single true Macrophage tumour.Five of the 11 patients with receptor-silent or immature B-cell tumours are alive between 7 and 15 months after the diagnosis. Diffuse mixed and diffuse poorly differentiated lymphocytic lymphomas in Rappaport's classification correlated poorly with receptors, mature and immature B-cell tumours being equally represented.
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PMCID: PMC2025534  PMID: 301396
22.  Cell receptor studies on seven cases of diffuse histiocytic malignant lymphoma (reticulum cell sarcoma). 
Journal of Clinical Pathology  1975;28(4):289-297.
Expression of B and T lymphocyte receptors has been studied in seven cases of reticulum cell sarcoma. In one case, surface receptors and tests of phagocytic function demonstrated the histiocytic origin of the neoplastic cells. In four cases, tumour cells expressed both B and T lymphocyte markers (two cases) or showed a normal pattern of expression of B and T lymphocyte markers. In the other two cases, lymphocyte receptors were not detected, and there was no evidence of phagocytic function: this class of receptor-silent tumours is of uncertain pathogenesis. The significance of these observations is discussed.
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PMCID: PMC475693  PMID: 1092721

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