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Immunoglobulin repertoire of B lymphocytes infiltrating breast medullary carcinoma.

Tumor specific peptides recognized by T lymphocytes infiltrating solid tumors, as well as the corresponding T cell receptor (TcR) repertoire usage, have been extensively investigated. By contrast, tumor infiltrating B cells and their immunoglobulin (Ig) repertoire have been studied only in a limited number of tumors. The objective of the present study was to determine, whether DNA sequence analysis of the expressed immunoglobulin variable regions of B cells that infiltrate breast cancer, could be used to reveal a potential specific tumor binding capacity of the antibodies. To answer this question, about 200 expressed Ig heavy (VH) and light chain variable gene (VL) regions were cloned, sequenced and comparatively analysed from a typical medullary beast carcinoma (MBC), where the massive B and plasma cell infiltration correlates with favourable prognosis despite of its high grade. The tumor infiltrating B cell Ig heavy and light chain sequences could be classified into clusters, families and subgroups, based on the identity level to germline, showing a pattern of oligoclonality. Some overrepresented clusters could be determined. In the course of a detailed analysis and search in Blastn database, a number of VH and VL sequences showed more than 99% homology to DNA sequences of Ig VH region, with proved tumor antigen binding capacity. Our data suggest, that potential tumor binder Ig VH and VL sequences might be selected using a detailed immunoglobulin variable region analysis. This new approach might have a benefit for further antibody engineering, as difficulties in search for tumor binders by phage library selection might be reduced and the time for selection shortened.

B-Lymphocytes↗

99mTc-interleukin-2 scintigraphy as a potential tool for evaluating tumor-infiltrating lymphocytes in melanoma lesions: a validation study.

UNLABELLED: Cutaneous melanoma is often characterized by the presence of tumor-infiltrating lymphocytes (TILs). The degree of such infiltration and cell activation are considered significant prognostic factors reflecting the host's immune response to the tumor; thus, patients with peritumoral infiltration may have a better prognosis and may also achieve a better response to interleukin-2 (IL2) immunotherapy. There is evidence that the expression of cluster designation (CD) 25 antigen (IL2 receptor [IL2R]) is a good marker of activity of T lymphocytes against melanoma cells. The aim of this study was to evaluate in vivo the binding of 99mTc-IL2 to lymphocytes infiltrating cutaneous melanoma and to determine whether such uptake correlates with immunologic and histologic data, thus providing useful prognostic information for IL2 therapy in patients with advanced disease. METHODS: Thirty patients with cutaneous lesions suspected of being melanoma were studied. Planar gamma-camera images over known tumor sites were acquired 1 h after the injection of 111-185 MBq of 99mTc-IL2. Tumor uptake of 99mTc-IL2 was measured as a target-to-background (T/B) radioactivity ratio. All patients underwent surgery, and histologic evaluation of the resected lesion was performed. The percentage of different peripheral blood lymphocyte subsets (CD3, CD4, CD8, CD16, CD25) and the percentage of IL2R-positive tumor cells on histologic sections were also measured. RESULTS: At final histology, 21 lesions were found to be melanoma and 9 were classified as benign. In 15 of 21 (71%) melanomas and 2 of 9 (22%) benign cutaneous lesions, we found uptake of 99mTc-IL2. The calculated T/B ratios correlated significantly with the number of IL2R-positive TILs. CONCLUSION: 99mTc-IL2 scintigraphy provides a means of in vivo measurement of the extent of tumor infiltration of IL2R-positive cells, thereby providing valuable prognostic information for selection of patients who may benefit from IL2 immunotherapy.

Feasibility Studies↗

[Fine needle aspiration cytology diagnosis of extramedullary leukemic infiltration].

OBJECTIVES: To assess the diagnostic accuracy of fine needle aspiration cytology (FNAC) in extramedullary leukemic infiltration. METHODS: The results of FNAC from 65 cases of extramedullary leukemic infiltration were reviewed and analyzed. RESULTS: In the 65 cases studied, there were 24 cases of acute lymphoblastic leukemia (ALL), 25 cases of acute myelogenous leukemia (AML), 6 cases of chronic lymphocytic leukemia (CLL) and 10 cases of chronic granulocytic leukemia (CML). The commonest site of infiltration was lymph node, which accounted for 73.8% of all cases. CONCLUSIONS: Accurate cytologic diagnosis of extramedullary leukemic infiltration relies on detailed morphologic assessment as well as correlation with clinical examination and other relevant laboratory findings, especially in patients whose initial symptom being a local mass. Leukemic infiltration, which represents proliferation of primitive cells, should be distinguished from non-Hodgkin's lymphoma. Morphologic assessment by oil immersion lens and examination of peripheral blood smears is useful in this respect.

Adolescent↗

Expression of the MAGE-A4 and NY-ESO-1 cancer-testis antigens and T cell infiltration in non-small cell lung carcinoma and their prognostic significance.

Cancer-testis (CT) antigens were identified as a group of highly attractive targets for cancer immunotherapy because of their expression in a variety of malignant tumors but solely in the testis among the normal adult tissues. To evaluate the potential of two members of this family, MAGE-A4 and NY-ESO-1 antigens, for cancer vaccine in non-small cell lung carcinoma (NSCLC), we examined the expression of these antigens and T cell infiltration in tumor tissue, and evaluated their prognostic significance. One hundred fifty-seven patients with NSCLC were studied. Reverse transcription-PCR was performed to evaluate MAGE-A4 and NY-ESO-1 expression. Immunohistochemistry was performed for NY-ESO-1 expression and T cell infiltration into the tumor site. Survival analysis was also performed. MAGE-A4 and NY-ESO-1 were expressed in 40 of 141 (28.4%) and 13 of 157 (8.3%) NSCLC respectively. Both CT antigens were more frequently expressed in squamous cell carcinoma (SCC) than in adenocarcinoma. An inverse correlation was found between MAGE-A4 expression and patient survival in advanced stage cancers. Combined infiltration of both CD4+ and CD8+ T cells into tumor nest predicted better survival. There was no correlation, however, between lymphocyte infiltration and antigen expression in the tumor. MAGE-A4 expression in advanced group and T cell infiltration may provide prognostic information. Lastly, these CT antigens, especially MAGE-A4, may represent potential targets for cancer immunotherapy in patients with NSCLC.

Adult↗

[Establishment of a nude mouse model of human monocytic leukemia with multi-organ infiltration].

BACKGROUND & OBJECTIVE: Acute leukemia is always accompanied by extramedullary infiltration, which may be the source of relapse. The mechanism of infiltration is unclear yet. This study was to establish a leukemic extramedullary infiltration model for exploring the mechanism. METHODS: After being treated by splenectomy, cytoxan intraperitoneal injection, and sublethal irradiation, 4-and 6-week old BALB/c nu/nu mice were transplanted intravenously with 5x10(6) or 1x10(7) of SHI-1 human monocytic leukemic cells. The leukemic cells engrafted into the mice were tracked by reverse transcription-polymerase chain reaction (RT-PCR), histopathologic examination, immunohistochemistry, and flow cytometry (FCM). RESULTS: SHI-1 cells were engrafted in nude mice and infiltrated into the stomach, liver, lungs, brain, kidneys, bone marrow, and so on, and formed green solid tumors in some positions. Autopsy revealed that neoplasms were found in more organs in 4-week old mice than in 6-week old mice. Human CD45-positive cells were detected in the bone marrow of 4-week old mice but not in 6-week old mice. CONCLUSION: SHI-1 cells can be engrafted in the pretreated nu/nu mice, and this model provides a useful experimental system for the research of leukemic extramedullary infiltration.

Animals↗

Methylcholanthrene-induced tumors in adult and in aging rats: infiltrating cells and their ADCC activity.

In our previous studies we found that the slower growth of syngeneic, immunogenic MC-induced sarcoma (MC-Sa) in aging rats was followed by the higher activity of spleen lymphocytes in ADCC assay. The purpose of the present paper was to study infiltrating cells of immunogenic and non-immunogenic MC-Sa and to estimate in situ ADCC activity in relation to the growth of MC-Sa transplants in adult and aging rats. It was found that MC-Sa's were infiltrated mainly by lymphocytes. Among cells infiltrating tumor the high percentage of cells with Fc receptor was present. During progressive tumor growth the percentage of infiltrating cells and also FcR+ cells significantly decreased within immunogenic MC-Sa, but did not change in the case of the non-immunogenic MC-Sa. In cell infiltrates of both tumors no differences between adult and aging rats were observed. At early stages of the tumor growth the cells active in ADCC assay were present within both MC-Sa. The slower growth of immunogenic MC-Sa in aging rats compared with adults was connected with longer maintenance of ADCC activity in situ. In the case of non-immunogenic MC-Sa, which grew at similar rate in both groups of rats, no differences in ADCC activity between adult and aging animals were observed. It is suggested that in situ ADCC activity may be one of the mechanisms responsible for the slower growth of immunogenic MC-Sa in aging rats.

Aging↗

Cytokine production and expression of adhesion molecules and integrins in tumor infiltrating lymphomononuclear cells of non-small cell carcinomas of the lung.

Localization of intercellular adhesion molecule-1 and vascular cell adhesion molecule-1, and of their ligands, lymphocyte function-associated antigen-1 and very late activation antigen-4, was determined in non-small cell lung carcinomas and tumor-free lung. Messenger RNA expression for interleukins (IL) IL-1 beta, IL-2, IL-4, IL-5, IL-6, IL-10, tumor necrosis factor-alpha, transforming growth factor-beta, interferon-gamma, granulocyte-macrophages colony stimulating factor, and human perforin-1 was assessed by in situ hybridization on the same tissues. Intercellular adhesion molecule-1 was expressed in all blood vessels, whereas only a low number of vessels displayed vascular cell adhesion molecule-1 immunoreactivity. Tumor infiltrating lymphomononuclear cells consisted of lymphocyte function-associated antigen-1-positive cells and of a lower number of very late activation antigen-4-positive cells. All squamous cell carcinomas consisted of intercellular adhesion molecule-1-positive neoplastic cells infiltrated by lymphocyte function-associated antigen-1-positive tumor infiltrating lymphomononuclear and CD-la-positive Langerhans cells, whereas only a minor number of adenocarcinomas displayed a consistent number of intercellular adhesion molecule-1-positive neoplastic cells. Tumor infiltrating lymphomononuclear cells showed a wider production of cytokines when compared to bronchus-associated lymphoid tissue of tumor-free lung. Moreover, cells producing interferon-gamma, IL-4, and IL-5 were more numerous in squamous cell carcinomas than in adenocarcinomas. These findings indicate that the lung squamous cell carcinoma might represent a neoplastic microenvironment able to induce activation of tumor infiltrating lymphomononuclear cells more efficiently than the adenocarcinoma.

Aged↗

Skin involvement in myelogenous leukemia: morphologic and immunophenotypic heterogeneity of skin infiltrates.

A systematic morphological analysis of cutaneous infiltrates in acute myelogenous leukemia and myelodysplastic syndrome revealed that in many cases the infiltrating cells have a different phenotype from those in the bone marrow. This study sought to answer two questions: (a) How wide is the range of cytological features and immunoreactivity of the cutaneous infiltrates and what danger is there of misinterpretation? (b) What are the possible causes of the wide spectrum of differentiation of the cells infiltrating the skin? Skin biopsy specimens from 16 patients with myelogenous leukemia or myelodysplastic syndrome were investigated. The diagnosis was acute myelomonocytic leukemia (M4, according to the French-American-British/FAB system of classification of acute leukemias) in eight cases, acute monocytic leukemia (M5) in four cases, aleukemic leukemia cutis as a recurrence of M2 leukemia after treatment in one case, and myelodysplastic syndrome in three cases, including one case of myelodysplasia with an excess of bone marrow blasts (RAEB-T) and two cases of chronic myelomonocytic leukemia, one of which presented as aleukemic leukemia cutis. Reactivity with the macrophage-associated antibodies anti-CD68, Ki-M1p, and anti-lysozyme was the most consistent. However, the naphthol AS-D chloroacetate esterase reaction and staining with DAKO-M1, Ki-My2p, anti-neutrophil elastase, and anti-CD34 were found to be of little value for identifying the cutaneous infiltrate as myelogenous. Some antibodies (e.g., anti-S100 protein and MB2) even produced staining in a few cases that could have led to a mistaken diagnosis of histiocytic neoplasm or malignant lymphoma.(ABSTRACT TRUNCATED AT 250 WORDS)

Antigen-Presenting Cells↗

Molecular diagnosis of lymphocytic infiltrates of the skin.

BACKGROUND: Advances in our understanding of the molecular genetics of lymphocyte antigen receptors (B-cell immunoglobulin and T-cell antigen receptor), have led to the application of molecular biologic techniques to molecularly characterize lymphocytic infiltrates of the skin. Molecular diagnosis refers to the application of these techniques as a diagnostic aid in the clinicopathologic evaluation of cutaneous lymphocytic infiltrates. OBSERVATION: Molecular studies have clinical application in the determination of lineage and detection of retroviruses in cutaneous lymphoid neoplasms, distinguishing between lymphoproliferative and reactive infiltrates, and staging and monitoring response to therapy in cutaneous T-cell lymphoma. Southern blot analysis of immunoglobulin and T-cell antigen receptor gene rearrangements may fail to aid the clinician in establishing a diagnosis of a cutaneous malignancy due to the limits of detection sensitivity in minimally infiltrated lesions (eg, parapsoriasis and patch-stage mycosis fungoides) or the still uncertain prognostic significance of clonality in benign cutaneous diseases (eg, follicular mucinosis, pityriasis lichenoides et varioliformis acuta, lymphomatoid papulosis, and cutaneous lymphoid hyperplasia). CONCLUSIONS: Molecular studies have enormous research value, providing new means to explore the pathogenesis and clonal evolution of lymphoproliferative skin diseases. Presently, however, they have limited applications as an independent diagnostic tool. As our understanding of the clinical and biologic significance of the molecular detection of clonal lymphocyte populations in the skin expands and as the application of polymerase chain reaction amplification provides us with greater detection sensitivity and specificity, the clinical utility of molecular diagnosis of lymphocytic infiltrates of the skin will be enhanced.

Antibodies, Monoclonal↗

[Histo-morphometry of bone mass and cellularity using image analysis: characterization of infiltration patterns in CLL].

PURPOSE: The aim of this study was to investigate a possible influence of lymphocytic infiltration in Chronic Lymphocytic Leukemia (CLL) on bone mass, according to the infiltration patterns. MATERIAL AND METHODS: We have studied histological preparations from decalcified and paraffin included bone biopsies and used a histomorphometric study through a mixed (interactive-automatic) image analysis, in order to quantify bone trabecular (BT/TV) and cellular volume (CV). Preparations were visualized via a 2.5 objective; a video camera was adapted and connected to a computer with analogical-digital converter incorporated. After the image was captured, a binarization was made by trabecular and medullary spaces segmentation; automatically, we calculated trabecular area (TA) and cellular area (CA). Expressions indicated above were used to calculate BV/TV and CV, expressed in percentage: BV/TV (%) = [TA/(TA + CA)] x 100, and CV (%) = [CA/(TA + CA)] x 100. RESULTS: 92 normal biopsies and 79 CLL biopsies were studied (Nodular 12, Diffuse 22, Interstitial 23, Mixed 22). BT/TV in normal subjects age over 45, was 18.40%; it was lower in CLL subjects, 17.25%. CV was 33.62% in normal cases and significantly higher in CLL with 42.95%. According to the infiltration patterns, Nodular pattern showed similar value to normal biopsies, versus Diffuse pattern with BT/TV of 16.19% and CV of 51.49%. Significantly, a high CV value was accompanied of a low B/TV value, in normal and CLL subjects. CONCLUSIONS: We analyzed histomorphometrically trabecular volume as expression of bone mass in undecalcified biopsies. CLL infiltration tended to show a lower trabecular volume. Diffuse pattern presented the higher cellular infiltration and bone mass destruction.

Aged↗

Correlation of apoptosis with the distribution and composition of lymphocytic infiltrate in human breast carcinomas.

We studied the lymphocytic infiltrate in human breast carcinomas and correlated its distribution and composition with the apoptotic index of cancerous cells. Paraffin sections from 36 human breast carcinomas with prominent lymphocytic infiltration were immunohistochemically stained with L26, UCHL1 and Leu-7 antibodies. Apoptotic index was expressed as the number of cells undergoing apoptosis per 20 HPF. The lymphocytic infiltrate was composed mainly of T-cells, usually peripherally but in 17 cases intratumorally distributed. T-cells, in contact with apoptotic cells, were observed in 11 cases. B-cells formed small aggregates at the periphery of the tumor. NK cells were present isolated in the infiltrate. No correlation was found between the type and distribution of the lymphocytic infiltrate and apoptotic index. The presence of intratumoral T-lymphocytes, occasionally observed in contact with apoptotic cells, may suggest that they could be partly associated with the apoptotic death of cancerous cells.

Apoptosis↗

Breast cancer: poor short-term prognosis in cases with moderate lymphocyte infiltration at the tumour edge: a preliminary report.

Lymphocyte infiltration in breast carcinoma has long been associated with favourable prognosis, however it has also been related to poor prognosis. In this study of 128 consecutive cases lymphocyte infiltration, its absence, or presence and degree was recorded at the tumour edge adjacent to fatty tissue in 100 cases, to other tissues in the remaining 28 in which no such edge was found. In the former there was a highly significant prognostic difference between those with moderate infiltration, which was of poor short-term prognosis, and those with other degrees of infiltration (none, slight and marked). Thus, lymphocyte infiltration in breast cancer can give information on both good and poor prognosis in over 80% of cases if its site and degree are taken into consideration. The remaining cases are histologically distinct. Their survival was similar to that in the total material. The mechanisms involved are as yet open to speculation. In view of increasing surgical reluctance to remove the axillary nodes it is of interest that the present investigation can be carried out on the routine diagnostic specimen from the primary and that the relation between survival and axillary nodal status, significant in the total material, no longer held in these groups.

Adenocarcinoma↗

Vascular adhesion protein-1 and ICAM-1 support the adhesion of tumor-infiltrating lymphocytes to tumor endothelium in human hepatocellular carcinoma.

T cell-mediated mechanisms are important in the defense against solid organ tumors. Why some tumors are more heavily infiltrated by T cells than others is poorly understood but is likely to depend upon adhesive interactions between circulating lymphocytes and tumor endothelium. In support of this hypothesis, the present study shows that primary human hepatocellular carcinomas (HCC) are more heavily infiltrated with T cells than colorectal hepatic metastases (CHM), and that their tumor vessels express high levels of several adhesion molecules. In HCC, an intense T cell infiltrate is observed within the tumor associated with strong expression of ICAM-1 and vascular adhesion protein-1 (VAP-1) on tumor endothelium. In contrast, fewer T cells infiltrated CHM and these tumors have little ICAM-1 and no detectable VAP-1 or VCAM-1 on tumor endothelium. T cells infiltrating both tumors are LFA-1 and very late Ag (VLA)-4 high. In vitro tissue-binding studies demonstrated that T cells bound readily to tumor endothelium in HCC, and Abs to ICAM-1, VAP-1, and to a lesser extent VCAM-1 could inhibit this binding. VAP-1 supported sialic acid-dependent adhesion under shear stress, suggesting that VAP-1 and ICAM-1 mediate, respectively, tethering and firm adhesion. In contrast, very few T cells bound to tumor vessels in CHM. Thus our data suggest that the VAP-1/VAP-1 receptor and ICAM-1/LFA-1 pathways are important in the recruitment of T cells to HCC. The strong expression of VAP-1 on tumor endothelium distinguishes HCC from CHM and supports our previous hypothesis that VAP-1 is an important hepatic endothelial adhesion molecule.

Adult↗

Lymphoid infiltration as a prognostic variable for early-onset breast carcinomas.

Infiltration by lymphoid cells is a common feature of many human tumors, including breast carcinomas, and the degree of infiltration has been suggested to be a measure of the host immune response. Our analyses in a series of 1919 cases of primary ductal and lobular infiltrating breast carcinomas from women with a long-term follow-up revealed: (a) a 16-17% frequency of infiltrated tumors independent of the patient's age at diagnosis; and (b) a strong positive correlation between survival rates and the presence of lymphocytes at the tumor site in patients less than 40 years of age (P = 0.0002) but no association with prognosis in patients 40 years of age or older. Multivariate analysis indicated that lymphoid infiltration is independent of other conventional prognostic factors such as nodal status and tumor size in predicting survival. Thus, a possible immune response against the tumor seems to be relevant only in women with early-onset tumors. Because the immune system is functionally maximum in younger years, declining with age, this finding might reflect a difference in the efficiency of the immune system. Alternatively, the biology of these tumors might differ, leading to a difference in immuno-genicity.

Adult↗

Pterygopalatine fossa and maxillary nerve infiltration in nasopharyngeal carcinoma.

BACKGROUND: Nasopharyngeal carcinoma (NPC) may infiltrate the pterygopalatine fossa (PPF) and the maxillary nerve. This study illustrates involvement of the maxillary nerve in the PPF with perineural spread to the cavernous sinus. METHODS: One hundred and fourteen patients with proven NPC were studied using magnetic resonance imaging (MRI) and computed tomography (CT). The images were retrospectively reviewed for PPF infiltration and maxillary nerve involvement. RESULTS: Seventeen (15%) patients showed infiltration of the PPF. Four patients had maxillary nerve involvement and a perineural spread to the cavernous sinus. Of the 17 patients with PPF infiltration, 8 (47%) patients showed hypoesthesia in the distribution of the infraorbital nerve. All 4 patients with contrast-enhancement of the maxillary nerve exhibited infraorbital neuropathy. CONCLUSION: Infiltration of the maxillary nerve in the PPF with intracranial spread is uncommon but should be suspected in patients with infraorbital neuropathy. This is important as it affects both prognosis and radiation treatment planning.

Cranial Nerve Neoplasms↗

Mononuclear phagocyte function in head and neck cancer. Chemotactic responsiveness of blood monocytes in correlation between histologic grade of the tumor and infiltration of these cells into the tumor area.

The chemotactic responsiveness of peripheral monocytes and the acid-phosphatase activity of tumor-infiltrating macrophages, as well as the ultrastructural appearance, were studied in 40 patients with squamous cell carcinoma of the head and neck. The chemotactic responsiveness was found to be decreased in carcinoma patients, and this value appeared to be positively correlated in individual patients with the number of tumor-infiltrating macrophages, as well as with the histologic grade of the tumor. Patients with poorly differentiated malignancies showed impaired monocyte chemotactic responsiveness and low numbers of tumor-infiltrating macrophages. Macrophages present in the parenchyma of the tumor showed a weak and diffuse pattern of acid phosphatase activity. The acid phosphatase activity of stromal macrophages was much stronger and distributed in foci. Electron microscopic examination of the parenchymal macrophages revealed low numbers of lysosomes and the presence of tumor cell debris in the cytoplasm of the cell, without any sign of a surrounding phagosomal membrane. Together with the weak cytochemical reactivity, this probably indicates the poor functional state of the phagocyte when infiltrated in the parenchyma of the tumor. Low molecular weight factors derived from the tumor are known to decrease chemotactic responsiveness of peripheral monocytes. The poor functional state of the macrophages infiltrated within tumor parenchyma might be explained by assuming that a high concentration of such factors in the near vicinity of malignant cells causes toxic effects in macrophages.

Adult↗

Splenectomy influences bone marrow infiltration in patients with splenic marginal zone cell lymphoma with or without villous lymphocytes.

BACKGROUND: Splenic marginal zone cell lymphoma (SMZL) is a low grade B-cell lymphoma in which patients can have circulating villous lymphocytes and can show a peculiar intrasinusoidal bone marrow (BM) infiltration. Splenectomy is the reported treatment of choice for these patients. The objective of this study was to evaluate the effects of splenectomy on patients with BM lymphomatous infiltration. METHODS: BM biopsies of 16 patients with SMZL were studied morphologically and immunohistochemically. In 12 patients, BM biopsies were taken before and after splenectomy. Four patients did not undergo splenectomy, and their BM biopsies were performed with an approximately 1 year interval. RESULTS: BM infiltration ranged from 10% to 40% of overall cellularity and was mostly of the intrasinusoidal type. After splenectomy, BM infiltration tended to become frankly nodular and showed an increase in tumor burden. Nonsplenectomized patients showed an unmodified picture. CONCLUSIONS: Splenectomy seems to induce important changes in patients with BM infiltration, probably through the lack of microenvironmental factors on circulating cells. These effects suggest reconsidering the role of splenectomy in the treatment of patients with SMZL.

Adult↗

Identification of T cells in early dermal lymphocytic infiltrates in avian scleroderma.

University of California, Davis (UCD) line 200 chickens spontaneously develop a progressive fibrotic syndrome with features similar to those observed in human autoimmune connective tissue diseases, including fibrosis, vascular occlusion, and lymphocytic infiltration of the comb, skin, digits, and viscera. Beginning at 2 weeks post hatch, line 200 chickens develop intense lymphocytic infiltration of the comb and dorsal neck skin. To further characterize the nature of these cellular infiltrates, weekly serial skin biopsy specimens from line 200 and control birds were examined using hematoxylin and eosin staining and indirect immunofluorescence with a library of mouse anti-chicken monoclonal antibodies specific for lymphocyte markers. In situ staining performed on serial skin sections revealed the presence of large groups of T cells beginning at 2 weeks of age. Further characterization of these infiltrates demonstrated the presence of both T helper and T cytotoxic/suppressor cells with a mean +/- SD T4:T8 ratio of 1.44 +/- 0.29 by 4 weeks of age. As the lesions progressed, the infiltrates also contained distinct groups of B cells as characterized by MUI 36. In addition, the lesions were strongly positive for B-L (Ia) antigen, which was noted on B cells, monocytes/macrophages, activated T cells, and fibroblasts. The skin sections were negative for 2 different macrophage monoclonal antibodies at all time-points. Upon extraction from affected skin, 42.0 +/- 13.06% (mean +/- SD) of these cells were positive for B-L, 35.10 +/- 6.51% were T cells, and 31.25 +/- 3.14% were recognized by MUI 36. Although positive staining for IgG was not found in these extracted cells, 7% of the isolated cells were positive for surface IgM.

Animals↗