局部物理和化学疗法对尖锐湿疣患者细胞免疫功能的影响

观察局部物理和化学疗法对尖锐湿疣病人机体免疫功能的影响。将45例尖锐湿疣病人分成两组 ,分别用0.5 %鬼臼毒素和高频电烧灼治疗 ,在治疗前后检测病人外周血NK细胞、T淋巴细胞亚群分布和白介素 2水平。结果 :尖锐湿疣病人的细胞免疫功能有异常。治疗前病人血中的CD8+细胞百分比显著增加 ,CD4+细胞百分比减少 ,CD4+/CD8+比率下降 ,血清白介素 2水平低于正常值 ,治疗后病人的细胞免疫功能有一定程度的恢复 ,但两组间的差异无显著性意义(P>0.05)。结果示局部物理或化学方法去除尖锐湿疣疣体对病人机体免疫功能的影响基本一致。     

EB virus-related angiocentric T-cell lymphoma

A 61-year-old woman developed an asymptomatic tumor on her right cheek. She had a history of surgical removal of a similar tumor on her left arm 2 years earlier. The facial lesions were flesh-colored, with 2×2 cm ulcers (Fig. 1). The left arm had ulcerative nodular lesions around the operation scar. Laboratory examinations revealed marked leukocytosis (19,510/μL) with lymphocytosis (54.6%). A few abnormal blastoid cells with large nuclei were detected by hemogram (Fig. 2). The red blood cell count, platelet count, coagulation tests, liver, and renal functions, including serum lysozyme, lipase, α1anti-trypsin, and angiotensin-converting enzyme activities, were normal, but a moderate elevation of thymidinekinase activity was observed. Anti-nuclear human T-cell leukemia virus type 1 (HTLV-1), human immunodeficiency virus (HIV), hepatitis B virus (HBV), and hepatitis C virus (HCV) antibodies were not detected. Mantoux reaction was negative and no DNA of tuberculosis was detected by polymerase chain reaction. Epstein–Barr virus (EBV) titers revealed EBV viral capsid antigen (VCA)-immunoglobulin G (IgG): ×160; VCA-IgM: undetectable level; early antigen (EA)-IgG: ×10; and EBNA: ×80. The lymphocytes showed normal proliferation responses to lectins. The tumor biopsy specimen revealed marked lymphocyte infiltration around vessels and focal necrosis in the deep dermis (Fig. 3a). The infiltrate was composed mainly of large blastoid lymphocytes with mitotic figures. Around the destroyed dermal vessels, tumor cells and denatured endothelial cells concentrated characteristically and formed glomeruloid lesions (Fig. 3b). Many auto-phagocytes forming “bean bag cells” were electron microscopically detected in the tumor. The proliferated cells in the dermis were CD3+, CD4+, CD7+, CD2+, CD5+, WT31+, and CD45RO+; however, CD1, CD8, TCRd1, CD16, CD56, CD57, CD19, CD20, CD10, L26, and LMP-1 were negative. Bone marrow cells showed lymphocytosis (39%) mixed with abnormal lymphoid cells. Rearrangement of T cell receptor (TCR)-β or TCR-γ chain was not detected in the samples from dermal tumors and peripheral blood. EBV-encoded small nuclear RNA (EBER) was detected in the tumor cells from the forearm by in situ hybridization. Thus, the patient was diagnosed as having EB virus-related angiocentric lymphoma with leukemic change, and was treated with bi-weekly cyclophosphamide, doxorubicin, vincristine, and prednisone (CHOP) combination chemotherapy. The facial tumor responded to CHOP markedly, and the peripheral blood leukemic cell count declined after two cycles of CHOP. After the sixth cycle of CHOP, however, the patient developed acute interstitial pneumonitis, and died from an intracranial hemorrhage. The autopsy specimens revealed thickening of the alveolar wall, septal fibrosis, and alveolar epithelial necrosis with focal lymphocyte infiltration. The bone marrow was hypocellular, but infiltrated with blastoid tumor cells.     

A rapid flow cytometric assay to detect CD4+ and CD8+ T-helper (Th) 0, Th1 and Th2 cells in whole blood and its application to study cytokine levels in atopic dermatitis before and after cyclosporin therapy

BACKGROUND: The immune response in atopic dermatitis (AD) is thought to be driven by T-helper (Th) 2 cytokines. Using flow cytometry, higher frequencies of peripheral blood CD4+ and CD8+ T cells producing interleukin (IL)-4 and correspondingly lower frequencies of CD4+ T cells producing interferon (IFN)-gamma have been found in patients with AD compared with healthy controls. It would be of interest to know whether other Th1 and Th2 cytokines such as IL-5, IL-13 and tumour necrosis factor (TNF)-alpha are similarly skewed in patients with AD and whether this immune skewing, detected via a simple blood assay, can be correlated with other clinical measurements or treatments in AD. OBJECTIVES: To use a rapid (4-h) flow cytometric assay to study a wide range of Th1 and Th2 cytokine patterns in peripheral blood lymphocytes from patients with AD, comparing them with non-atopic healthy controls. To correlate cytokine patterns with the degree of eosinophilia observed and in the case of one patient with severe disease, to observe the effect of cyclosporin therapy on peripheral blood cytokine patterns. METHODS: Peripheral blood from eight patients with AD and 23 healthy controls was examined for the frequencies of CD4+ and CD8+ T cells expressing IL-2, IL-4, IL-5, IL-13, IFN-gamma and TNF-alpha using flow cytometry. RESULTS: Significantly higher frequencies of CD4+/IL-4+ (P < 0.005) and CD4+/IL-13+ (P < 0.0001) and lower frequencies of CD4+/IFN-gamma+ (P < 0.002) and CD8+/TNF-alpha+ (P < 0.05) T lymphocytes were found in patients with AD compared with controls. There were significant positive correlations with the increased percentages of CD4+/IL-4+ and CD4+/IL-13+ T lymphocytes and the degree of eosinophilia observed (P < 0.05, P < 0.001) and a negative correlation between the percentage of CD4+/IFN-gamma+ T lymphocytes and eosinophilia (P < 0.05). In one patient examined before and 8 days after cyclosporin therapy, 50% or greater reductions were observed in percentages of peripheral blood CD8+/IL-5+, CD8+/IL-13+, CD4+/IL-4+ and CD4+/IL-5+ T lymphocytes following cyclosporin therapy. A smaller reduction of 15% after cyclosporin therapy was found in percentages of CD4+/IL-13+ T cells. CONCLUSIONS: These data strongly support a Th2 predominance in the peripheral blood of AD. The results suggest that administration of cyclosporin therapy in patients with AD may help to restore the Th2 cytokine imbalance seen in these patients.     

Erythema multiforme-like lesions associated with lesional infiltration of tumor cells occurring with adult T-cell lymphoma/leukemia

A 66‐year‐old Japanese woman visited our hospital with a complaint of multiple papules on her trunk and extremities. She had a past medical history of appendicitis and blood transfusion 40 years earlier. For the last 10 years, she had noticed multiple, gradually enlarging papulonodular lesions with surrounding erythema on her trunk and extremities. Physical examination revealed multiple, violaceous papules or nodules, less than 10 mm in diameter, with surrounding erythema on her trunk and extremities ( Fig. 1 ). The results of routine laboratory examinations, including blood count, liver function, renal function, serum calcium, and lactate dehydrogenase, were within the normal range. The peripheral blood picture showed a small population of atypical lymphocytes below 1% of the total white blood cells. Human T‐cell lymphotropic virus type I (HTLV‐I) serology was positive. A microscopic examination of a biopsy specimen from a nodule on the abdomen demonstrated diffuse infiltration of large pleomorphic T cells in the upper and middle dermis, although highly atypical lymphocytes, so‐called flower cells, could not be recognized. Infiltrating lymphocytes were positive for CD2, CD3, CD4, CD5, CD7, and CD45, but negative for CD8 and CD20, immunohistologically. Bone marrow biopsy also demonstrated the infiltration of lymphocytes expressing CD2, CD3, CD4, CD5, and CD7, but not CD25. Southern blot analysis of the infiltrating cells in the skin revealed an integration of HTLV‐I proviral DNA in T cells. Clonal T‐cell receptor γ gene rearrangement was detected in skin and bone marrow biopsies. No abnormal mass or bone defect was detected by chest or abdominal computed tomographic scanning, systemic gallium‐67 citrate scintigraphy, or chest radiography. On the basis of these data, the patient was diagnosed with smouldering‐type adult T‐cell lymphoma/leukemia.

Figure 1 Open in figure viewer PowerPoint Clinical features of adult T‐cell lymphoma/leukemia (ATL) skin lesions. Crusted, target‐like, dark‐red plaques on the lower legs     

The diagnosis of Sézary syndrome on peripheral blood by flow cytometry requires the use of multiple markers

Background Diagnosis of Sézary syndrome (SS)‐defining blood involvement is hampered by the lack of Sézary cell‐specific markers and nonspecific morphology of the tumour cells. Objectives To identify the most reliable and easy to use markers for the diagnosis of SS‐defining blood involvement. Methods We studied 17 patients with SS and 11 control patients. We used flow cytometry for the detection of T‐cell antigens (CD3, CD4, CD7 and CD8), expression of the Sézary cell‐associated marker CD158k and T‐cell receptor (TCR)‐Vβ chain. Additionally, Sézary cells were identified by peripheral blood smear for lymphocytes with cerebriform nuclei. Results It was not possible to diagnose blood involvement in all patients with SS by a single marker or method, although none of the markers was increased in the control population. Sézary cells were detected by blood smears in 13 of 17 (76%), by flow cytometry by their CD4+ CD7? CD3^dim phenotype (> 1000 cells μL^?1) in 13 of 17 (76%) and by expression of CD158k in 11 of 17 (65%) patients with SS. A specific T‐cell clone was identified by identical TCR‐Vβ chain expression in 12 of 17 (71%) patients with SS. The identification of Sézary cells in individual patients varied for the different markers investigated. Conclusions The combination of identifying CD4+ CD7? CD3^dim cells, TCR‐Vβ chain and CD158k expression allowed a definite identification of SS‐defining blood involvement in every individual patient. All of these markers can be measured by flow cytometry which would avoid time‐consuming analysis of blood smears. These markers would also be suitable to monitor tumour cell load during therapy.     

The relevance of the CD4+ CD26- subset in the identification of circulating Sézary cells

BACKGROUND: The lack of specific markers for the phenotyping of circulating neoplastic T cells in Sézary syndrome (SS) patients makes it difficult both to ascertain the presence of clonal cells and to quantify the tumour burden in the peripheral blood. In previous reports we showed that the lack of CD26 (dipeptidyl-aminopeptidase IV) is a characteristic feature of circulating Sézary cells (SC). OBJECTIVES: The purpose of this study was to ascertain, by means of high-resolution two-, three- or four-parameter flow cytometry, the relationship between CD26 expression on peripheral blood lymphocytes and peripheral blood involvement in cutaneous T-cell lymphoma patients and to assess its significance in SS diagnosis. METHODS: The patient population included 52 SS patients, 151 mycosis fungoides (MF) patients at different clinical stages (including 14 with blood involvement, B1-MF), 88 patients with erythrodermic inflammatory skin diseases (EISD) and 72 healthy donors (HD). CD26+ values were available in all cases, whereas CD4+ CD26- level measurement was performed in 23 SS, 141 MF, 71 EISD and 72 HD. RESULTS: CD4+ CD26- percentage values were higher than 30% in all but one B1-MF and higher than 40% in all SS cases, whereas HD, EISD and B0-MF patient values were always lower than 30%. A statistically significant difference was found in both CD26- and CD4+ CD26- percentage and absolute values between SS and HD, EISD and B0-MF patients. The CD26- and CD4+ CD26- percentage values (but not the absolute values) were significantly higher in B1-MF compared with HD, EISD and B0-MF patients (P < 0.001). Moreover, CD26- absolute values and CD4+ CD26- percentage and absolute values were significantly higher in SS than in B1-MF (P < 0.001). A statistically significant direct relationship was found between CD4+ CD26- percentage values and the percentage of circulating SC within the lymphoid population in SS and B1-MF (r = 0.77; P < 0.001). The lack of CD26 was confirmed on phenotypically clonal cells in patients with an expanded circulating TCRvbeta population or a T-cell antigen loss. Sorted CD4+ CD26- cells from both SS patients and HD showed the characteristic cerebriform nuclei of SC. CONCLUSIONS: We feel that a CD4+ CD26- percentage value higher than 30% of peripheral blood lymphocytes could correctly identify the presence of peripheral blood involvement in SS and MF patients.     

RPR持续阳性梅毒患者脑脊液梅毒抗体与外周血细胞免疫的相关性研究

目的检测经过治疗但RPR持续阳性两年以上无神经损害临床表现的梅毒患者脑脊液中梅毒抗体与外周血淋巴细胞亚群,并探讨其相关性.方法应用梅毒血清学试验等方法对46例HIV阴性的这类梅毒患者的脑脊液进行梅毒抗体检测;同时应用流式细胞仪检测这些患者外周血淋巴细胞亚群,并与5例未治神经梅毒的脑脊液进行梅毒抗体检测结果和30例健康人群的外周血检测结果相对照.结果46例患者脑脊液中,有12例(27.39%)存在梅毒抗体,5例神经梅毒的脑脊液均存在梅毒抗体.患者外周血CD3 、CD4 T细胞及自然杀伤细胞与健康人群的检测结果差异无显著性(P＞0.05),而CD8 T细胞明显高于对照组,差异有显著性(P＜0.001);脑脊液梅毒抗体阳性和阴性梅毒患者之间,外周血淋巴细胞亚群检测结果差异无显著性(P＞0.05).结论经过治疗但RPR持续阳性梅毒患者脑脊液存在梅毒抗体,表明部分患者存在无症状神经梅毒;经过常规治疗但RPR持续阳性梅毒患者存在细胞免疫缺陷;无症状神经梅毒与外周血细胞免疫异常无明显的相关性.     

Papuloerythroderma‐like cutaneous involvement of a CD62L− subclone of T‐cell prolymphocytic leukemia

We report the case of an 88‐year‐old Japanese man with erythrodermic involvement of T‐cell prolymphocytic leukemia (T‐PLL). He had a history of pharyngeal diffuse large B‐cell lymphoma successfully treated with polychemotherapy including cyclophosphamide and epirubicin, 6 years before the current illness. He presented with numerous reddish, coalescing, flat‐topped papules on the trunk and extremities, sparing the skin folds of the abdomen, the features of which mimicked those of papuloerythroderma. Immunohistochemistry showed perivascular and epidermotropic infiltration of CD3⁺ CD4⁺ T cells in the cutaneous lesion. However, flow cytometric analysis revealed that the skin infiltrating T cells were negative for surface CD4, and that CD3⁺ CD4^? CD8^? cells made up 92% of the T‐cell fraction of peripheral blood. The circulating atypical T cells had a round or oval nucleus and prominent nucleoli, and the deletion of chromosomes 6q, 13 and 17. These cytological profiles were consistent with those of T‐PLL and distinct from those of Sézary cells. The same T‐cell clone was detected in the cutaneous lesion and peripheral blood, but the expression of CD62L was absent in the skin infiltrates and present in the circulating cells. No specific mutation was detected in STAT3 or STAT5B. Although low‐dose oral etoposide had a beneficial effect on the skin rash, a fatal crisis of marked leukocytosis (169 × 10³/μL) occurred 19 months after the illness onset. CD62L‐leukemic cells of T‐PLL may infiltrate the skin to form papuloerythroderma‐like cutaneous lesions.     

CD4+ CD8+ (thymocyte-like) T lymphocytes present in blood and skin from patients with atopic dermatitis suggest immune dysregulation

BACKGROUND: Atopic dermatitis (AD) is a chronic inflammatory skin disease expressed early in life. Disease development is primarily determined by as yet unknown genetic factors, leading to the accumulation of activated T lymphocytes in the skin. OBJECTIVES: To investigate the nature of these T cells. METHODS: T-cell lines could be established from AD skin biopsies, but not from normal skin or AD peripheral blood, when placed in RPMI 1640 medium with 10% human AB serum, antibiotics, and the T-lymphocyte growth factors interleukins 2 and 4. The cell lines were subjected to phenotypic analysis using a fluorescence-activated cell sorter and compared with lymphocytes from AD and normal control peripheral blood. RESULTS: T-cell lines from 22 of 24 consecutive skin biopsies taken from 24 adult patients with AD were established. All cells were T lymphocytes expressing several activation markers. A significant proportion of the lymphocytes had stable expression of a CD4+ CD8+ phenotype (26% +/- 6%; mean +/- SEM). Such double-positive T lymphocytes are normally only seen in the thymus and not in the peripheral immune system. CD4+ CD8+ cells in peripheral blood of the patients (12.5% +/- 3.3%) were also detected. CONCLUSIONS: We suggest that a basic pathophysiological change in AD may be a faulty maturation of the T-lymphocyte system, leading to skin inflammation with CD4+ CD8+ T lymphocytes resembling immature T cells. This is likely to lead to skewing of many immune reactions in the patients.     

Multiple cutaneous reticulohistiocytosis with T‐cell large granular lymphocyte clonopathy

A 63‐year‐old Caucasian man presented with a 4‐month history of disseminated asymptomatic reddish‐brown papulonodular lesions. A skin biopsy showed dermal infiltration with CD68⁺ histiocytes, predominantly with eosinophilic cytoplasm, some with a ground‐glass cytoplasm, and a small number of giant cells. The diagnosis of multiple cutaneous reticulohistiocytosis was made. Bone marrow immunophenotyping due to peripheral blood lymphocytosis revealed the presence of a monoclonal population of CD3⁺, CD8⁺ CD57⁺ large granular lymphocytes. The present case suggests the coexistence of multiple cutaneous reticulohistiocytosis with an underlying disorder.     

Increased expression of a novel early activation surface membrane receptor in cutaneous T cell lymphoma cells

Using a newly generated monoclonal antibody we identified the 96 kDa transmembrane receptor SC5 expressed simultaneously on a human Sezary cell line and a minor T cell subset in normal individuals. SC5 antigen was detected mostly on CD45RO+ lymphocytes from both CD4+ and CD8+ subsets as well as on natural killer and B lineage cells. SC5 surface expression increased very early after polyclonal stimulation of CD3+ cells due to the transfer of intracellular SC5 molecules to the cell membrane. Engagement of SC5 receptor by its monoclonal antibody inhibited the anti-CD3-induced proliferation and cytokine secretion of peripheral blood T cells and cell clones, whereas SC5 monoclonal antibody did not affect the cytotoxic activity of CD8+ T cell clones. Extensive phenotypic analysis revealed that the percentage of SC5+ CD4+ circulating lymphocytes in Sezary syndrome patients was significantly increased in comparison with controls (p < 0.01) and correlated with the morphologically detected percentage of Sezary syndrome cells in peripheral blood (p < 0.001). In one patient we clearly demonstrated that the circulating malignant T cells coexpress SC5 molecules. Importantly, ligation of SC5 receptor in a cutaneous T cell lymphoma cell line profoundly inhibited the anti-CD3-induced proliferation. Consequently, the expression of SC5 receptor in the peripheral blood of Sezary syndrome patients may serve not only to detect the presence of circulating malignant CD4+ cells but also as a target for immunotherapy.     

Two Cases of Metastatic Malignant Melanoma of the Lower Limb Treated with Hyperthermic Isolated Limb Perfusion and Concomitant Infusion of Either Carboplatin or β-Interferon

Two cases of metastatic malignant melanoma of the lower limb who were treated successfully with hyperthermic isolated limb perfusion are reported. One patient was infused with cis-diammine (1.1-cyclobutanedicarboxylate) platinum (II) (carboplatin, Paraplatin®, Bristol-Myers Squibb Company, New Jersey, USA), and the other was infused with human natural β-interferon (Feron®, Toray, Tokyo, Japan), via the external iliac artery. The first case showed a remarkable suppression of the growth of multiple metastatic melanoma nodules associated with numerous melanophage infiltrations, as shown histopathologically after the operation. The patient's serum level of 5-S-cysteinyl dopa decreased for the two months following the treatment In the second case, new formation of metastatic melanoma nodules was completely suppressed for up to 12 months following the operation. Analysis of immunological parameters showed that the number of peripheral CD8⁺ lymphocytes gradually and constantly increased after the operation, while that of CD4⁺ lymphocytes transiently increased and then returned to the pre-operative level. Natural killer activity transiently decreased to a slight degree 4 days after the operation and then returned to the pre-operative level 21 days after the operation. Side effects, such as nausea, vomiting and leg discomfort, were seen in the patient (Case 1) treated with carboplatin, but were completely reversible. These results suggest that hyperthermic isolated limb perfusion with concomitant infusion of carboplatin or β-interferon is effective in suppressing the growth of metastatic malignant melanomas of the lower limb.     

Eosinophile Zellulitis (Wells-Syndrom)

Eosinophilic cellulitis (Wells' syndrome) is a rare disorder characterized clinically by recurrent erythematous plaques resembling cellulitis and histologically by a dermal infiltrate of lymphocytes, eosinophils and eosinophil debris between collagen bundles, forming flame figures in typical cases. A 71-year-old woman with Wells' syndrome with blood and bone marrow eosinophilia showed a good response to dapsone. The level of eosinophil cationic protein (ECP) in serum was elevated. Immunophenotyping of peripheral T cells revealed an increased proportion of CD3+CD4+T cells. The patients' cultured peripheral lymphocytes spontaneously released significant amounts of interleukin 5 (IL-5), but not interleukin 4 (IL-4) or interferon gamma (IFN gamma). These findings suggest that activated T cells may be involved in the pathogenesis of blood and tissue eosinophilia in this patient.     

CUTANEOUS B-CELL LYMPHOMA CONSISTING OF LARGE CLEAVED CELLS WITH MULTILOBATED NUCLEI

A 65-year-old man was seen at the Ushioda Hospital in Au-gust 1989, because of a 1-month history of a tumor on the scalp. The tumor was excised and the diagnosis was malig-nant lymphoma. The patient was then referred to our de-partment in September 1989. Several nut-sized lymph nodes wvepa-b and m-fepa for 2 months. Since then, the patient has been free of disease up to the time of writing, July 1992, a period of 2.5 years. Biopsy samples taken from the tumor on the scalp showed a monomorphous infiltrate of large lymphoid cells throughout the entire dermis and subcutis, with a definite clear zone (Fig.1). A high-power view showed diffuse large lymphoid cell infiltration. Numerous mitotic figures were also seen. The lymphoid cells had multilobated nuclei and distinct nucleoli (Fig. 2). Monoclonal antibodies such as Leui (CD5), Leu2a (CD8), Leu3a (CD4), Leu4 (CD3), MT-1 (CD43), Leu14 (CD22), LN1 (CDw75), and Leu26 (CD20), and polyclonal antibodies such as anti-kappa, anti-lambda, anti-IgG, anti-lgA, anti-IgM, and anti-lgD were purchased from commercial sources. Optimal dilutions of the monoclonal antibodies and heteroantisera were assessed beforehand by titration on suitable tissue samples. The antigens recognized by the monoclonal anti-bodies and heteroantisera were investigated by either the avidin-biotin peroxidase complex (ABC) method on cryostat sections or the peroxidase-antiperoxidase complex (PAP) method on paraffin sections, as described elsewhere.¹ The immunologic properties of the infiltrating cells were determined using skin biopsied in August 1989, and October 1989. Large lymphoid cells, which formed the major popu-lation of infiltrating cells, were positive for CD20, CD22, and HLA-DR and negative for CD3, CD4, CD43, and CD45RO. From these findings the patient was diagnosed as hav-ing primary cutaneous B-cell lymphoma, diffuse large non-cleaved cell type, as classified by the Working Formulation.²     

Granuloma annulare associated with Hodgkin's disease

A 39-year-old man was referred to our institution for treatment of Hodgkin's disease (HD) previously diagnosed at another hospital. On physical examination the patient had bilateral cervical lymphadenopathy (9 cm to 0.5 cm in size), but no clinical hepatosplenomegaly was noted. Examination of his skin showed indurated reddish macules and papules scattered over the trunk and buttocks (Fig. 1). Laboratory examination showed a hemoglobin of 13.9 g/100 mL, with a leukocyte count of 8600/mm³ with 69% neutrophils, 9% eosinophils, 2% monocytes, 22% lymphocytes, and 2% abnormal lymphocytes. The platelet count was 398 × 10³/mm³. The serum IgG was 1877 g/100 mL, the IgA was 572.5 g/100 mL, and the IgM was 55.5 g/100 mL, Serum test for HTLV-1 antibody was positive, but hepatitis B antigen and hepatitis C antibodies were negative, A ⁶⁷Ga scintigram showed uptake in the supraclavicular and right retrocervical areas. Computed tomographic scans showed bilateral cervical lymph node and left axillary lymph node enlargement. Bone marrow aspiration was normal. A lymph node biopsy specimen from the left axillary region showed partially effaced lymph node architecture with a few immature or atypical mononuclear cells and some Reed-Stemberg cells. Necrosis and fibrosis was not seen (Fig. 2), The diagnosis of HD, lymphocyte predominant subtype, was therefore confirmed and he was staged as HD-IIA. Biopsies were also obtained from the skin of his back and abdominal wall which showed perifollicular and perivascular cellular infiltration composed of mature small lymphocytes, epithelioid histiocytes, and multinucleated giant cells. Neither Hodgkin cells nor Reed-Sternberg cells were recognized in the lesional skin. Collagen bundles lying in the upper dermis and close to the adipose tissue showed focal necrobiotic change particularly in the center of the infiltration (Fig. 3). Alcian-blue stain confirmed acid mucopolysuccharide deposition on and between degenerated collagen bundles. Special stains for fungus and mycobacterium were negative. Immunohistochemically, the epithelioid histiocytes and multinucleated giant cells labelled positive for HLA-DR in the membrane and were positive for lysozyme and alpha-1 antichymotrypsin in the cytoplasm. Intermingled with these cells, S100a positive dendritic cells were also present, especially around hair follicle. The lymphocytes were immunoreactive with UCHL-1 (CD45RO), Leu3 (CD4), and Leu4 (CD3), However, only a few lymphocytes stained with Leu3a (CD8), These findings of the skin lesions were consistent with those of granuloma annulare (GA). The patient was started on combination chemotherapy ABVD (adriamycin, bleomycin, vinblastin, and DTIC) for the HD, After 10 courses of the chemotherapy, the patient is disease free. The GA lesions also began to disappear following the institution of the chemotherapy and the lesions were mostly gone after a second course of ABVD.     

Selective generation of CD8+ T-cell clones from the peripheral blood of patients with cutaneous reactions to beta-lactam antibiotics

The presence, phenotype, and functional characteristics of peripheral blood penicillin-specific T lymphocytes in individuals with cutaneous allergic reactions to penicillin were investigated using in vitro long-term culture techniques. Peripheral blood mononuclear cells from two penicillin-allergic patients were stimulated in vitro with penicillin, and T-cell blasts were clonally expanded by limiting dilution. Seven T-cell clones were derived, all of which were CD3⁺ CD4^? CD8⁺ HLA-DR⁺, and produced IL-2 and IFN-γ upon stimulation. T-cell proliferation required the presence of antigen and autologous, but not allogeneic, antigen-presenting cells. In addition to the parent compound, the T-cell clones also developed a proliferative response to penicilloyl, the major metabolite of penicillin. The cloned T-cell lines were found to exhibit marked suppressor activity for Con A mitogenesis. The observed suppressor activity required cell-to-cell contact, as supernatants from these T-cell clones had no comparable inhibitory effect. These findings indicate that there is a predominance of penicillin-specific CD8⁺ T cells in the peripheral blood of individuals sensitized to beta-lactam antibiotics.     

CD4/CD8 double-positive, acute type of adult T-cell leukemia/lymphoma with extensive cutaneous involvement

A 67‐year‐old Korean man presented with a 2‐week history of pruritic cutaneous lesions on the trunk and a 2‐month history of cervical neck masses. The cutaneous lesions had been spreading rapidly for the last week. The past history included an unknown liver disease about 40 years previously, and he had never been transfused. The physical examination revealed multiple cervical lymph node enlargements and a two‐finger width of splenomegaly with tenderness. Numerous nodules and plaques were distributed on the trunk, face, and proximal extremities ( Fig. 1 ). The results of laboratory tests were as follows: leukocyte count, 24,200/mm³, with lymphocytes 15,560/mm³; serum lactate dehydrogenase, 943 IU/L (normal range, 263–450 IU/L); alkaline phosphatase, 135 IU/L (normal range, 35–60 IU/L); blood urea nitrogen, 32 mg/dL (normal range, 3–24 mg/dL); serum creatinine, 1.7 mg/dL (normal range, 0.3–1.6 mg/dL); serum calcium, 17.5 mg/dL (normal range, 8.8–10.2 mg/dL). Other laboratory results, including liver function test and urine analysis, showed no abnormalities. The examination of peripheral blood revealed multilobulated atypical lymphoid cells. The radiologic images showed hilar, para‐aortic lymph node enlargement and splenomegaly. A skin biopsy from the nodular lesion revealed massive infiltration of small‐ to medium‐sized atypical lymphoid cells in the dermis ( Fig. 2a ). The infiltrating cells were positive for CD3 ( Fig. 2b ), CD4 ( Fig. 2c ), CD5, and CD8 ( Fig. 2d ), but negative for CD20, CD34, CD56, CD68, and terminal deoxynucleotidyl transferase (TdT) in the immunohistochemical studies on paraffin sections. They also showed high Ki‐67 labeling (80–90%), and this finding reflects their highly proliferative nature. Polymerase chain reaction (PCR) analysis showed a distinct band for the T‐cell γ receptor gene, confirming the T‐cell clonality of the infiltrating neoplastic cells. Specimens from the cervical lymph node and bone marrow showed the same results in immunohistochemical studies. PCR analysis of the DNA from peripheral leukemic cells showed human T‐cell leukemia virus type I (HTLV‐I) proviral integration. The patient was diagnosed as having the acute type of adult T‐cell leukemia/lymphoma (ATLL). The disease course was extremely aggressive, and he died of acute renal failure on the 16th day following diagnosis.

Figure 1 Open in figure viewer PowerPoint Pruritic erythematous nodules and plaques on the face, trunk, and upper arms     

Sézary syndrome presenting with 'leonine facies'

A 71-year-old man presented with erythroderma and multiple nodular skin lesions over the face, scalp, upper limbs and trunk. The facial skin was thickened, producing the rare 'leonine facies' appearance. Investigations revealed the presence of atypical lymphoid cells in the peripheral blood, bone marrow and skin. The atypical lymphoid cells in the peripheral blood and bone marrow were positive for helper T-cell antigens (CD4, CD2, CD5 and CD7) on immunophenotyping by flow cytometry. The histopathology of skin showed dermal infiltration by atypical small lymphocytes with epidermotropism. These cells were positive for helper T-lymphocyte antigens on immunohistochemistry. A diagnosis of Sézary syndrome was made based on clinical, peripheral blood and immunophenotypical findings.     

T cells migrate to tumour sites after extracorporeal interleukin 2 stimulation and reinfusion in a patient with metastatic melanoma

Peripheral blood mononuclear cells (PBMC) were taken by leukapheresis from a patient with melanoma skin metastases and stimulated in vitro using 1000 IU recombinant interleukin 2 (IL-2)/ml to generate lymphokine-activated killer cells (LAK cells). Two-colour immunofluorescence analysis demonstrated an IL-2-induced up-regulation of CD25 on natural killer cells (CD56⁺) as well as on T lymphocytes (CD3⁺). After radiolabelling with indium-111, the cells were reinfuse. Gamma-camera imaging revealed an enrichment at the tumour sites. Immunostaining of tumour tissue taken before and after scintigraphy demonstrated CD25⁺ Tlymphocytes (CD2⁺, CD3⁺), but no natural killer cells (CD16⁺, CD56⁺) infiltrating the metastases. LAK cell enrichment at melanoma metastases in vivo did not involve natural killer cells, but was characterized by increased numbers of activated T lymphocytes in this patient.