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PubMed1997

Immunophenotypic analysis of ovarian and testicular Müllerian papillary serous tumors.

Intratesticular Müllerian papillary serous tumors lacking stromal invasion are uncommon neoplasms whose immunophenotypic properties have not been studied extensively. We present such information here and compare it with information from a group of ovarian papillary serous tumors of low malignant potential ("borderline serous tumors") that are morphologically identical. We compared the histologic features of our index case of intratesticular Müllerian papillary serous tumor with those of nine ovarian papillary serous tumors. We then evaluated both the index case and the ovarian tumors with antibodies against carcinoembryonic antigen, LeuM1, CA125, estrogen receptors, progesterone receptors, cytokeratin 7, and cytokeratin 20, by use of established immunohistochemical techniques. The testicular and ovarian tumors were morphologically indistinguishable. The intratesticular Müllerian papillary serous tumor expressed LeuM1, CA125, estrogen receptors, progesterone receptors, cytokeratin 7, and weak cytokeratin 20; carcinoembryonic antigen was not expressed. All of the ovarian papillary serous tumors expressed CA125, estrogen receptors, and cytokeratin 7. Eight of nine expressed progesterone receptors. Five of nine stained with LeuM1. Two of nine were focally weakly positive with cytokeratin 20. LeuM1 expression helps distinguish testicular papillary serous tumors from mesothelial proliferations, which might seem morphologically similar. The immunophenotype of intratesticular and female genital papillary serous tumors is similar; this similarity extends to expression of estrogen and progesterone receptors, which is rare in neoplasms in men, especially among testicular neoplasms.

Aged↗

PubMed1997

Evolution of leukocyte immunophenotyping as influenced by the HIV/AIDS pandemic: a short history of the development of gating strategies for CD4+ T-cell enumeration.

This brief review of the development of gating strategies for CD4+ T-cell enumeration is really the story of contemporary clinical flow cytometry. It is the chronicle of its birth, and its slow invasion into the clinical immunology laboratory over the past 15 years. The driving force behind the technological evolution of leukocyte immunophenotyping was, and still is, the impact of the HIV/AIDS pandemic on the discipline of immunology. While, at times, this review of analytical flow cytometry makes metaphorical excursions into the literary world, the objective is not to trivialize technology. Rather, it attempts to illustrate how, in his drive to solve the practical problems of modern medicine, man's essentially eclectic nature excels in an environment of diverse knowledge. This review will examine how technological advancements, such as the introduction of clinical instruments with multi-color capabilities, and the evolution of leukocyte gating strategies, has influenced the evolution of leukocyte immunophenotyping.

Acquired Immunodeficiency Syndrome↗

PubMed1997

Classical Hodgkin's disease. Clinical impact of the immunophenotype.

Antibodies against CD15, -30, and -20 are often used to support morphological diagnosis of Hodgkin's Disease (HD). The classical HD, i.e., the non-lymphocyte-predominance types, are CD15+, CD30+, and CD20- in general. However, the results for CD15 are less clear-cut in many studies, showing up to 40% of classical HD that lack positivity for this maker. Little is currently known about the relevance of antigen expression in relation to clinical outcome in HD. Therefore, the three markers were analyzed in 1751 cases from the German Hodgkin Study Group, using micro-wave epitope retrieval to optimize staining sensitivity. Eighty-three percent of the cases showed a classical immunophenotype (CD15+, CD30+, CD20-), twelve percent lacked CD15 positivity (CD15-, CD30+, CD20-), and five percent showed other combinations. For 1286 cases, clinical follow-up was available, which revealed significant differences for freedom from treatment failure (P = 0.0022) and overall survival (P = 0.0001) between cases with classical immunophenotype and CD15 negativity (CD30+, CD20-). Multivariate Cox regression using the three markers, age, sex, histology, stage, B-symptoms (fever, sweats, weight loss > 10% of body weight), hemoglobin, and erythrocyte sedimentation rate as factors showed that lack of CD15 expression in classical HD is an independent negative prognostic factor for relapses (P = 0.022) and survival (P = 0.0035). In conclusion, immunohistochemistry is able to identify classical HD cases with unfavorable clinical outcome.

Adolescent↗

PubMed1997

Immunophenotyping as a diagnostic tool to differentiate lichen planus from chronic graft-versus-host disease: diagnostic observations on two patients.

BACKGROUND: Lichen planus (LP) and the lichenoid variant of chronic graft-versus-host disease (cGVHD) can present with similar clinical and histological findings. The distinction, although difficult, is important both prognostically and therapeutically. The mechanism and effector cell phenotypes have also shown to differ between the 2 entities. While the lichenoid infiltrate of LP is predominantly T lymphocytes helper/inducer cell phenotype, the suppressor/cytotoxic subset appears to play a major role in cGVHD. The aim of this study is to determine whether the immunophenotypic character of the lichenoid infiltrate can aid in distinguishing the 2 entities. METHODS: Biopsies were obtained from 2 patients with lichenoid papules and a history of transplantation. Light microscopy revealed lichenoid inflammation in both cases characterized by a band-like lymphohistiocytic infiltrate at the dermal-epidermal junction. Immunochemistry was performed on fresh tissue using a panel of monoclonal antibodies including anti-CD1a, CD3, CD4, CD8, CD16, CD20, CD28, and CD68. Results were quantitated using computer-assisted image analysis. RESULTS: We found that in both cases the majority of cells stained with pan T cell marker CD3+. One case demonstrated predominantly CD4+ T cells and increased numbers of CD1a positive Langerhans cells, while the lymphokine natural killer cell activity (LAK) markers anti-CD16 and anti-CD28 were largely nonreactive. Conversely, the second case contained predominately CD8+ lymphocytes and very few CD1a positive Langerhans cells with abundant LAK cell anti-CD16 and anti-CD28 reactivity. CONCLUSIONS: Based on these findings, the former was classified as lichen planus and the latter as lichenoid cGVHD. The diagnoses are substantiated with clinical history and follow-up information. We conclude that immunophenotypic characteristics of the infiltrate can be a useful tool in differentiating lichenoid cGVHD from lichen planus.

Adult↗

PubMed1998

Immunophenotypic characterization of simian immunodeficiency virus-infected dendritic cells in cervix, vagina, and draining lymph nodes of rhesus monkeys.

Significant progress has been made in understanding the biology of heterosexual transmission of HIV by utilizing the simian immunodeficiency virus (SIV)/rhesus monkey animal model. Our previous studies have shown that SIV-infected cells within the stratified squamous epithelium of the vagina have a dendritic morphology. However, the type of cell infected was not conclusively determined. The purpose of the present study was to immunophenotype the SIV-infected cells in the lower reproductive tract and genital lymph nodes of the female rhesus monkey. Vagina, cervix, and iliac lymph node from eight chronically SIV-infected adult female monkeys were examined for this study. None of the animals had histologic evidence of opportunistic infections or genital tract pathogens other than SIV. Combined in situ hybridization and immunohistochemistry were used to detect SIV RNA and to determine the immunophenotype of SIV-infected cells in tissue sections and cytospin preparations of cells from the tissues. We now show that SIV-infected cells were most common in iliac lymph node and that the majority of infected cells in the lymph node were T lymphocytes. SIV-infected macrophages, Langerhans' cells, and dendritic cells were also found in the lymph node. SIV-infected cells were found within the epithelium and lamina propria of the vagina. Although most of the infected cells were T cells, a significant proportion (approximately 40%) of the SIV-infected cells in cytospin preparations from explant cultures of vagina and cervix were Langerhans' cells. SIV-infected T cells in the lower genital tract were commonly associated with focal mononuclear cell infiltrates. SIV-infected macrophages were rarely found in the genital tract. The present study provides the first direct demonstration that Langerhans' cells and dendritic cells in the genital tract and lymph nodes are infected with SIV in vivo. Thus, dendritic cells, in general, and Langerhans' cells, in particular, are important reservoirs for HIV/SIV replication in vivo.

Animals↗

PubMed1998

[Assessment of degree of malignancy in non-Hodgkin B-cell lymphoma with immunophenotyping].

Due to immunophenotypic examination of bioptic samples from 30 patients with non-Hodgkin's lymphoma (NHL), malignancy of B-cell NHL was identified at an early stage of diagnosis, before histological analysis yielded the results. Negative expression of T-cell antigens, monoclonal character of light-weight chains of immunoglobulins, positive expression of B-cell antigens and marked expression of CD5+ are immunologic markers of low-malignancy B-cell NHL. A similar immunophenotypic pattern, involving negative expression of CD5- and positive one for Calla-antigen as well as one identifiable by monoclonal non-cluster antibody IPO3, is a marker of highly-malignant B-cell NHL.

Antibodies, Monoclonal↗

PubMed1998

Immunophenotypic characterization of plasma cells from monoclonal gammopathy of undetermined significance patients. Implications for the differential diagnosis between MGUS and multiple myeloma.

Although the immunophenotype of plasma cells (PCs) from multiple myeloma (MM) patients has been extensively explored, information on the phenotypic characteristics of PCs in monoclonal gammopathy of undetermined significance (MGUS) patients is scanty and frequently controversial. Thus, the question of whether or not PCs are phenotypically different in the two disorders and whether this criteria could be useful for the differential diagnosis between MGUS and MM remains to be explored. In the present study, the immunophenotypic profile of bone marrow PCs (BMPCs) from a group of 76 MGUS patients has been analyzed by flow cytometry and compared with that of BMPCs present in both MM patients (n = 65) and control subjects (n = 10). For that purpose, a large panel of monoclonal antibodies against PC-related antigens was used together with a sensitive methodology in which a minimum of 10(3) PCs were studied. In all MGUS cases studied, two clearly defined and distinct PC subpopulations could be identified. One PC subpopulation, population A (33 +/- 31% of total PCs), constantly displayed a high CD38 expression with low forward light scatter (FSC)/side light scatter (SSC) and was positive for CD19 and negative for CD56 (only a small proportion of these PCs were weakly positive for CD56). The other PC subpopulation, population B (67 +/- 31% of total PCs), showed the opposite pattern; the antigen CD56 was strongly positive and CD19 was constantly negative, and it showed a lower CD38 expression and higher FSC/SSC values than population A. Clonality studies (cytoplasmic light chain restriction, DNA content studies, and polymerase chain reaction assessment) confirmed the clonal nature of PCs from population B and the polyclonal origin of PCs from population A. Moreover, the polyclonal PCs from MGUS displayed a phenotypic profile identical to that found in PCs from healthy individuals. By contrast, clonal PCs from all MGUS patients displayed a similar antigenic profile to myelomatous PCs, with clear phenotypic differences with respect to normal PCs: lower intensity of CD38 expression and a variable reactivity for markers that were not expressed in normal PCs, such as CD28, CD117, and sIg. Although the presence of residual polyclonal PCs was a constant finding in MGUS patients, it was a rare event in MM and, when present (only 22% of MM cases), its frequency was significantly lower than that observed in MGUS (0.25% versus 32.9%, respectively; P < 0.0001). Only 1.5% of patients with MM had more than 3% of normal PCs, whereas 98% of patients with MGUS had more than 3%. Moreover, as shown by multivariate analysis, the number of residual polyclonal PCs was the most powerful single parameter for the discrimination between MGUS and MM patients at diagnosis, even when only stage I MM cases were considered.

Adult↗

PubMedDate not supplied

Fetal cells in maternal blood: isolation by magnetic cell sorting and confirmation by immunophenotyping and FISH.

Fetal cells entering the maternal circulation during pregnancy constitute a potential source for safe and reliable non invasive prenatal diagnosis. However, selecting the appropriate fetal cell type and methods of enrichment are areas of paramount importance. Most investigators consider fetal nucleated red blood cells (NRBCs) to be the cell type of choice, since they are mononuclear, abundant in fetal blood, relatively well differentiated and have a limited life span. Twenty ml of peripheral blood samples were collected from 40 pregnant women in the 16th to 18th week of pregnancy. To enrich for NRBCs, found within an excess of maternal cells, negative magnetic cell sorting (MACS) was used. Leukocytes were depleted from maternal blood by treatment with anti CD45 monoclonal antibody, as this surface antigen is not expressed in NRBCs. NRBCs were detected in 35 of the 40 maternal samples with May Grunwald-Giemsa staining. In 30 cases UCH gamma positive cells were identified after immunophenotyping with a monoclonal antibody directed against the gamma chain of fetal hemoglobin. The mean number of isolated NRBCs was 6 (range 1-15). In 5 cases we were able to successfully perform FISH on the immunophenotyped cells and determine correctly the fetal gender using X and Y chromosome specific probes.

DNA Probes↗

PubMed1997

[Immunophenotypic and clinical features of Philadelphia chromosome positive adult lymphocytic leukemia].

OBJECTIVE: To sutdy the immunophenotypic and clinical features of Philadelphia chromosome positive adult acute lymphocytic leukemia (Ph+ ALL). METHODS: Fifty-one previously untreated Ph+ ALL were included. Morphologic/cytochemistric, cytogenetic and immunophenotypic examinations were performed. Patients received daunorubicin-vincristine-prednisone (DOP) or DOP-asparaginas (DOLP) induction protocols. Two patients received autologous bone marrow transplantation (ABMT) as postremission therapy. RESULTS: The 51 patients were among 158 patients with ALL (32.3%). Patients with Ph+ ALL were older and had a significantly higher incidence of peripheral leukocytosis and blasts, FAB L2 subtype, B lineage antigens expression and CD34 positivity. 42.1% of Ph+ ALL achieved complete remission (CR). Their remission duration was 4.5 months, the median time to CR was 54 days. Two patients receiving ABMT survived in remission 22 and 33 months, respectively. CONCLUSION: Patients with Ph+ ALL are usually older, have FAB L2 morphology, and are B lineage antigens, CD34 positive. These patients had lower CR rate, shorter remission duration, and needed longer time to achieve CR.

Adult↗

PubMedDate not supplied

Immunophenotype, ras oncogenes and p53 onco-suppressor gene in benzo(a)pyrene induced malignant soft tissue tumours in Wistar rats.

This study was designed to identify the immunophenotypic characteristics of malignant soft tissue tumours, induced experimentally with benzo(a)pyrene (BaP), and to evaluate the immunohistochemical expression of the ras oncogene family and p53 onco-suppressor gene in these tumours, in association with prognostic factors. Seventy-five male Wistar rats were subcutaneous injected, dorsally, with a single dose of 10.08 mgr BaP. A solid, well-circumscribed tumour was formed at the injection site, in 70 of the animals, 80-100 days after the carcinogen's administration. The tumour as well as selected main organs were excised and studied after the animals' death. All the specimens were fixed in formalin 10%, embedded in paraffin and stained with H + E. The immunohistochemical avidin-biotin method was performed in the tumour sections, using the following monoclonal or polyclonal antibodies: vimentin, desmin, muscle specific actin (MSA), a-smooth muscle actin (SMA), myoglobin, smooth muscle myosin, a-1-antitrypsin, a-1-antichymotrypsin, S-100 protein, epithelial membrane antigen (EMA), K-ras, H-ras, Pan-ras and p53. The induced tumours of the animals were almost well-circumscribed, with a partly storiform cut surface. Histologically, their appearance was more conventional with high grade leiomyosarcomas; about half of them showed highly anaplastic areas, resembling other pleomorphic undifferentiated sarcomas. Pulmonary metastatic foci were detected in 37 animals. Immunohistochemically, all the tumours displayed positive expression of vimentin, MSA and SMA. Desmin was positively expressed in 40 tumours, smooth muscle myosin in 57 tumours and EMA in 12 tumours. All the tumours were negative for myoglobin, a-1-antitrypsin, a-1-antichymotrypsin and S-100 protein. In addition, five tumours showed a positive reaction for K-ras p21, 37 for H-ras p21, 41 for Pan-ras p21 and 14 for p53 protein. The overexpression of the oncoproteins H-ras p21 and Pan-ras p21 in these tumours was significantly associated with a non-advanced tumour stage (absence of metastatic focus). In conclusion, the histological as well as the immunophenotypic features of the induced tumours are more conventional with leiomyosarcomas mostly of high grade; many of them are "dedifferentiated". The identification of both ras and p53 gene products in these tumours indicates that alterations of these genes are common but not specific events, implicated in the tumourigenesis, which may become prognostic markers for this subtype of soft tissue sarcomas.

Animals↗

PubMed1998

[Multi-parametric analysis for leukemic immunophenotyping].

We have investigated whether immunophenotyping measured by laser flow cytometry could be corresponding to FAB classification of acute leukemias, using multi-parametric analysis. Sixty-one patients with acute leukemias have been evaluated, including 23 patients with acute lymphocytic leukemias (ALL) and 38 patients with acute myelogenous leukemias. In principal component analysis, positive cell surface antigens could be classified into groups along to the cell lineage and differentiation. In discriminant analysis, the sensitivity by immunophenotypic method to FAB subtypes was 75%, whereas the specificity was over 90%. In cluster analysis, patients have been classified into 4 groups, which essentially corresponded to ALL, M1/M2, M3 and M4/M5. Based on those multi-parametric analysis, a new flow chart has been established, resulting that the sensitivity and the specificity was improved to over 90% and 95% respectively. These results suggest that the classification of acute leukemia using the flow chart could be useful tools for diagnosis of subtypes of acute leukemias.

Antigens, Surface↗

PubMed1999

Primary plasma cell leukemia: clinical, immunophenotypic, DNA ploidy, and cytogenetic characteristics.

We report on a series of 26 patients diagnosed with primary (de novo) plasma cell (PC) leukemia (PCL) in whom we analyzed the clinicobiologic characteristics of the disease together with the immunophenotype, DNA cell content, proliferative index, and numeric chromosomal aberrations of the neoplastic PC, and compared them with 664 multiple myeloma (MM) patients at diagnosis. The median age, sex ratio, and bone lesion extension were similar, but PCL cases displayed a higher prevalence of clinical stage III, extramedullary involvement, and Bence Jones cases, with fewer IgA cases than for MM patients. In addition, according to several prognostic indicators (beta2-microglobulin serum level, proportion of S-phase PCs, proteinuria, calcium serum level, lactate dehydrogenase [LDH] and renal function), the incidence of adverse prognostic factors was significantly higher in PCL versus MM. Immunophenotypic expression was similar for CD38, CD138, CD2, CD3, CD16, CD10, CD13, and CD15, but PCL differed from MM in the expression of CD56, CD9 HLA-DR, CD117, and CD20 antigens. Twenty-two PCL cases were diploid and one was hypodiploid, while most MM cases (57%) showed DNA hyperdiploidy. With the fluorescent in situ hydridization (FISH) technique, 12 of 13 PCL cases displayed the numeric aberrations, -13 (86%), +/-1 (57%), +18 (43%), and -X in women (25%), but they lacked several numeric aberrations usually found in MM such as +3, +6, +9, +11, and +15. PCL cases had a lower overall response to therapy than MM cases (38% v 63%, P =.01332). Among PCL patients, a trend for a worse response was observed in cases treated with melphalan and prednisone (MP) versus polychemotherapy. Overall survival was significantly worse in PCL versus MM patients (8 v 36 months, P <.0001), but it was significantly better in PCL patients treated with polychemotherapy versus MP (18 v 3 months, P =.0137). By contrast, MM patients did not show significant differences in overall survival according to the treatment used, MP or polychemotherapy. Ten variables seemed to predict survival in PCL patients, but only the beta2-microglobulin level and S-phase PCs retained an independent value in multivariate analysis. In summary, our study illustrates that PCs from PCL display singular phenotypic, DNA cell content, and cytogenetic characteristics that lead to a different disease evolution versus MM.

Aged↗

PubMed1987

A clinicopathologic and immunohistochemical study of 16 patients with small intestinal leiomyosarcoma. Limited utility of immunophenotyping.

Clinicopathologic data including immunophenotypic expression of 16 new cases of small intestinal leiomyosarcoma are presented. Patient age ranged from 27 to 87 years (mean, 61 years) with a 2.2:1 male-female ratio. Eighty-eight percent of tumors occurred distally, i.e., in the jejunoileum. Considerable histologic heterogeneity was noted including epithelioid, myxoid, and sclerotic variants. Stratification by histologic grade suggests important clinical subsets. Only in grade I tumors is surgical extirpation alone potentially curative. Grade II tumors are associated with meaningful survival usually measured in years, however, nearly all patients can be expected to die from their tumors. Grade III sarcomas are fully malignant and rapidly progressive: except for one case with limited follow-up, all patients with Grade III sarcomas died of disease with a mean survival of only nine months (median, 2 months). Immunostaining with antibodies directed against vimentin, muscle-specific actin, desmin, and S-100 protein were performed. Only antivimentin was uniformly reactive. Sixty-three percent of cases tested showed actin immunoreactivity, and no tumors revealed immunohistochemically detectable desmin or S-100 protein. Although many small bowel stromal sarcomas appear "poorly differentiated" by immunohistochemical methods presently available, that they are indeed leiomyosarcomas seems most likely from their intramural location and light histologic study. The limited utility of immunophenotyping and the importance of histologic grading are stressed.

Actins↗

PubMed1988

Immunophenotypic differences between plasmacytoma/multiple myeloma and immunoblastic lymphoma.

Four cases of plasmacytoma (PC), six cases of multiple myeloma (MM), and nine cases of immunoblastic lymphoma (IL) of B-cell phenotype were studied with a large panel of monoclonal antibodies applied to frozen tissue sections. There were no significant differences in the immunophenotypes of plasmacytomas and multiple myelomas. However, significant immunophenotypic differences were noticed between the plasmacytoma/multiple myeloma cases (PC/MM) and the immunoblastic lymphoma specimens. The PC/MM cases characteristically stained with alpha (or gamma) and T10 and did not usually stain with mu, leukocyte common antibodies, certain B-lineage antibodies (B1, T015, 4G7, 6A4), or Ia. In contrast, IL sections usually did not stain with alpha or T10 and generally did stain with mu (or gamma), leukocyte common antibodies, B-lineage antibodies, and Ia. Ki-67, an antibody to proliferating cells, stained significantly fewer cells in PC/MM than in IL and stained significantly fewer cells that had a good clinical outcome. We conclude that although no one antibody is useful in distinguishing PC/MM from IL, the application of a panel of antibodies may be helpful in making this distinction. The prognosis may correlate with the numbers of proliferating cells as measured by reactivity with Ki-67.

Antibodies, Monoclonal↗

PubMed1989

Immunophenotypic spectrum of plasma cell leukemia.

Four cases of plasma cell leukemia (PCL) are reported that illustrate the variable immunotype of this disorder in contrast with the immunologic profile described for normal B-cell maturation and typical multiple myeloma (MM). Mature B-lymphocytes express B1 antigen (Ag) and surface immunoglobulin (SIg) whereas maturation to the plasma cell stage is accompanied by loss of these immunologic markers and expression of T10 Ag, plasma cell Ag (PCA), and cytoplasmic immunoglobulin (CIg). Plasma cells from patients with MM have previously been found to express the immunophenotype of normal plasma cells. In contrast, none of the four cases reported here express an immunologic profile typical for specific B-cell differentiation stages. Only one of four cases was strongly positive for PCA but additionally expressed B1 Ag and SIg. Of the remaining three cases, all expressed T10 Ag and CIg; two cases also expressed SIg, weak PCA, and B1 Ag. All four cases were monoclonal for lambda light chains and negative for common ALL Ag (CALLA). The variable expression of mature B-cell markers and plasma cell markers demonstrates the immunophenotypic spectrum of PCL; the prognostic significance of this heterogeneity needs to be more closely examined.

Adult↗

PubMed1989

Immunophenotyping in the classification of acute leukemia in adults. Interpretation of multiple lineage reactivity.

Fifty-nine adult patients with acute leukemia were classified using a combination of the French-American-British (FAB) criteria and characterization by immunophenotyping using flow cytometric study. The authors identified 51 patients with acute myeloblastic leukemia and eight with acute lymphoblastic leukemia. This procedure permitted lineage assignment in leukemias that otherwise might have been unclassifiable. In addition, the authors demonstrated that the leukemic blasts of 29% of patients with myeloblastic disease exhibited one or more T-cell antigens on their surface. The use of immunophenotyping has greatly enhanced the authors' ability to correctly identify the lineage of acute leukemias. The data, however, must be interpreted with caution with respect to diagnosing acute mixed lineage leukemias and must be integrated with the morphologic and cytochemical evaluation of traditional classification schemes. The possible significance of T-cell markers in myeloblastic leukemia is discussed.

Adult↗

PubMed1989

Comprehensive quality assessment approach for flow cytometric immunophenotyping of human lymphocytes.

Flow cytometric immunophenotypic (IPT) evaluation has become an important adjunct to clinical patient management and epidemiological studies. This has precipitated a need for stringent quality assessment (QA) procedures to ascertain data integrity. We evaluated a QA approach to monitor all elements of the immunophenotyping process, inclusive of blood collection and processing procedures as well as of staining reagent and instrument performance. Central to our approach was preparation each day, in parallel with clinical analytes, of lymphocytes from healthy donors, selected from a 15 donor panel. IPT parameters evaluated over a 19 month period included frequencies of CD3+, CD4+, CD8+, and CD20+ lymphocytes and the ratio of CD4+ to CD8+ lymphocytes. The sensitivity for analytical error detection was reflected by median coefficients of variation of these parameters within individual panel donors, which were 4.1%, 4.5%, 3.9%, 8.2%, and 10.1%, respectively. IPT parameter values were determined each day for two of the panel donors, then averaged and standardized to obtain a quality or Q variate, which was the basis of QA. Error detection sensitivity decreased 0.6-1.7% and the number of false rejections increased 1.2-3.3% when one panel donor rather than two was used daily for QA. This study also illuminated important aspects of what constitutes the norm for longitudinal IPT parameter variation in healthy individuals including: 1) a generally low degree of temporal parameter variation within individual donors, but 2) significant differences between donors with respect to variance estimates for CD3+ and CD8+ lymphocyte frequencies and CD4+/CD8+ lymphocyte ratios, and 3) an apparent seasonal pattern of variation in CD4+ T-cell frequencies.

Antibodies, Monoclonal↗

PubMed1987

Clonal rearrangements of T-cell receptor and immunoglobulin genes and immunophenotypic antigen expression in different subclasses of Hodgkin's disease.

Twenty-two cases of Hodgkin's disease (HD), representing the 4 different subclasses, were studied by immunophenotypic and immunogenotypic analysis. Quantitative immunophenotypic analysis of HD infiltrates showed a predominance of CD3-positive T cells in all subtypes except the lymphocytic depletion (HDLD) subtype. Only 5 samples of HD [2 of lymphocytic predominance (HDLP), 2 of mixed cellularity (HDMC), and one of nodular sclerosis type (HDNS)] were found to have both their Ig and T-cell antigen receptor (TcR) genes in the germ-line configuration. The remaining patients with HDLP (3 cases), HDNS (5 cases), and HDMC (4 cases), all exhibited rearrangements of either TcR gamma or TCR gamma and TcR beta genes, while all 5 cases of HDLD had either TCR gamma or immunoglobulin heavy-chain gene rearrangement. These results substantiate the view that Hodgkin's lymphomas contain clonal lymphocyte populations and that different rearrangement patterns may be associated with different subclasses of HD.

Antigens, Differentiation, T-Lymphocyte↗