Complete molecular remission during biologic response modifier therapy for Sézary syndrome is associated with enhanced helper T type 1 cytokine production and natural killer cell activity.

BACKGROUND: Cutaneous T-cell lymphoma (CTCL) is a clonally derived, skin-invasive malignancy of CD4(+) T lymphocytes with the phenotype of mature helper T cells. Advancing stages of CTCL are associated with depressed cell-mediated immunity, increased production of T helper type 2 cytokines and decreased levels of T helper type 1 cytokines. OBJECTIVE: Our purpose was to evaluate the cytokine secretion pattern and cell-mediated cytotoxicity in peripheral blood mononuclear cells of patients with Sézary syndrome in relation to the presence of the malignant clone. METHODS: Serial polymerase chain reaction for the T-cell receptor-beta or T-cell receptor-gamma gene rearrangement was used to determine the presence of the malignant clone. Enzyme-linked immunosorbent assays were used to determine the levels of interleukin 4 and interferon gamma produced by the peripheral blood mononuclear cells from the patients with Sézary syndrome. RESULTS: We demonstrate 3 cases of Sézary syndrome with typically suppressed cell-mediated cytotoxicity, elevated production of interleukin 4, and depressed production of interferon gamma by their peripheral blood mononuclear cells before institution of therapy with biologic response modifier therapy. In all 3 cases after clinical and molecular remission, we observed striking immunologic changes, including an increase in levels of natural killer cell activity and interferon gamma production and decreased production of interleukin 4. CONCLUSIONS: The observation that the cytokine secretion pattern by peripheral blood mononuclear cells from 3 patients with Sézary syndrome normalized with the disappearance of the malignant clone from the peripheral blood suggests that the malignant T cells account for the aberrant cytokine production. Moreover, the aberrant cytokine production may be the cause for suppression of cell-mediated immunity seen in advancing stages of CTCL.     

Natural cell-mediated cytotoxicity against various target cells in patients with epidermodysplasia verruciformis

Natural cell-mediated cytotoxicity of peripheral blood mononuclear cells was studied in eight patients with epidermodysplasia verruciformis induced by human papillomaviruses specific for epidermodysplasia verruciformis and in five patients with epidermodysplasia verruciformis-induced exclusively by human papillomavirus type 3. Nine patients with various cutaneous warts and 25 age- and sex-matched healthy persons were control subjects. Natural cell-mediated cytotoxicity against both K-562 erythroleukemic and Sk-v cells was in the normal range in patients with epidermodysplasia verruciformis induced by epidermodysplasia verruciformis-specific human papillomaviruses and in patients with cutaneous warts. The lysis of both targets, however, was significantly decreased in patients with the form of epidermodysplasia verruciformis associated with human papillomavirus type 3. Experiments with normal keratinocytes and with keratinocytes isolated from a malignant lesion bearing human papillomavirus type 5 genomes showed that the latter were susceptible to lysis by the peripheral blood mononuclear cells of healthy persons and of patients with cutaneous warts. Lysis of keratinocytes in epidermodysplasia verruciformis, however, was strongly reduced in patients with epidermodysplasia verruciformis induced by specific human papillomaviruses. This reduction was not associated with a decrease in anti-K-562 natural cell-mediated cytotoxicity. Our results suggest that in patients with epidermodysplasia verruciformis induced by disease-specific human papillomaviruses, there is reduced natural cell-mediated cytotoxicity against epidermodysplasia verruciformis keratinocytes.     

自体外周造血干细胞移植治疗成人皮肌炎一例

目的 探讨自体外周造血干细胞移植治疗成人皮肌炎的疗效.方法 1例21岁皮肌炎患者接受自体外周造血干细胞移植,随访6年.采用重组人粒细胞集落刺激因子动员外周造血干细胞后,予以环磷酰胺、甲泼尼龙、环孢素预处理,干细胞回输第3天开始使用兔抗人T淋巴细胞免疫球蛋白.观察症状体征、生化指标、造血重建及免疫恢复情况.结果 出院时皮疹明显好转,后逐渐消退;四肢肌力由移植前Ⅳ级转为Ⅴ级并持续至今;移植后肌酸激酶显著下降,但又逐渐恢复到移植前水平;回输当天开始出现白细胞下降,第8天恢复正常造血功能.移植前后免疫功能均正常.结论 对复杂难治性重症皮肌炎,可选择白体外周造血干细胞移植治疗.     

Stimulation of T cells by autologous mononuclear leukocytes and epidermal cells in psoriasis

Summary Based on reports suggesting aberrant cell-mediated immunity and altered infiltration of immunocompetent cells into the skin in psoriasis, we studied the stimulation of T cells by autologous non-T mononuclear leukocytes (autologous mixed lymphocyte reaction, AMLR) and by epidermal cells isolated from lesional and clinically uninvolved skin in psoriasis (autologous mixed epidermal cell lymphocyte reaction, AMECLR). Age- and sex-matched individuals served as controls. We found that the AMLR in psoriasis (n=11) was similar to that in healthy controls (n=16); furthermore, cell proliferation was alike in the presence of either 5% AB-serum or autologous serum. By contrast, while the AMECLR in healthy controls (n=9) resembled that in psoriatics employing epidermal cells from univolved skin, epidermal cells from lesional sites (n=10) induced a significantly higher proliferation of autologous T cells in the AMECLR (P<0.01). We conclude that the in vitro stimulation of T cells by non-T mononuclear leukocytes is normal in psoriasis and is not regulated by autologous serum. Lesional psoriatic epidermal cells, however, are more active in stimulating autologous T cell proliferation than cells from univolved psoriatic or normal epidermis.     

Lymphocyte responsiveness to urinary glycosaminoglycan in systemic scleroderma.

From the urine of patients with systemic scleroderma, we previously isolated a glycosaminoglycan, which could induce a scleroderma-like change in the skin of mouse. In the present work, the cell-mediated response to urinary glycosaminoglycans was examined by lymphocyte transformation test. The specific response was observed in lymphocytes of patients with scleroderma, when the above scleroderma-inducing glycosaminoglycan was added. By contrast, lymphocytes of patients with SLE or dermatomyositis and of normal persons hardly showed a response to this glycosaminoglycan. On the other hand, the glycosaminoglycans from normal urine could not stimulate lymphocytes of patients with scleroderma or healthy persons.     

Immunoregulatory properties of serum from patients with different stages of syphilis

The response of lymphocytes from 17 patients with primary, secondary, and tertiary syphilis to phytohaemagglutinin (PHA) and to allogeneic lymphocytes was normal in heterologous serum; however, the responsiveness of cells from some patients with primary and secondary disease was significantly reduced in the presence of autologous serum. As cells from healthy controls invariably responded better to these stimuli in autologous serum, the sera from 81 patients with syphilis were screened for immunosuppressive properties. Sera from 25 primary, 32 secondary, two tertiary, six congenital, and 16 latent cases of syphilis were examined for their ability to reduce the responsiveness of normal cells to PHA. These experiments were performed with test sera as the sole source of serum for the cultures or with test sera added to cultures containing optimally supportive amounts of pooled human plasma.

Stimulation of normal cells from one control in human plasma and 20% test serum showed that only in sera from congenital cases of syphilis was the mean response significantly different from the response seen in control sera; a significant increase in the response to stimulation occurred. The range of response to PHA with sera from cases of primary and secondary syphilis was wider than with normal sera. Sera from five (20%) cases of primary and 14 (44%) cases of secondary syphilis appeared to be immunosuppressive. When retested on another sample of normal cells, these sera were consistently immunosuppressive even in the presence of 15% pooled human plasma. Thus, in early syphilis antigenic stimulation may result in the release from suppressor cells of non-specific immunoregulators of cell-mediated immunity. Such phenomena may be a prelude to the development of tolerance to treponemal antigens. In congenital syphilis the development of suppressor cells may be impaired, resulting in the apparent immunostimulatory properties of serum from such cases.

     

Lymphokine-activated killer (LAK) activity against autologous malignant tumors of the skin

When peripheral blood mononuclear cells (PBMC) cultured with interleukin 2 (IL-2) develop the ability to lyse fresh tumor cells, such activity is referred to as lymphokine-activated killer (LAK) activity. In this paper, we examined LAK activity against the autologous skin tumors, malignant melanoma (MM), squamous cell carcinoma (SCC), and basal cell epithelioma (BCE), which have distinct clinical characteristics. Similar levels of LAK activity against Daudi 1A4, an NK resistant cell line were significantly obtained from all cancer patients. However, different levels of LAK activity against autologous tumor cells were found from three kinds of cancer patients using mixtures of autologous tumors and LAK. The levels of cytotoxicity were 29.8 +/- 7.0% in five MM, 15.9 +/- 4.9% in seven SCC, and 4.0 +/- 2.3% in five BCE patients at an effector/target ratio of 50/1. Allogeneic MM targets were lysed by LAK from all three types of cancer patients at similar levels, implying that LAK is not restricted to major histocompatibility complex (MHC) antigens. These results indicate that the levels of autologous LAK activity were significantly associated with the magnitude of clinical malignancy of the tumor targets, and suggested the selective lysis of tumor targets by LAK. NK activity of cancer patients bearing tumors was also examined prior to therapy. The levels of NK activity observed in MM patients were considerably lower than those in two other cancer patients.     

Immune sensitization against epidermal antigens in polymorphous light eruption

To get further insight into the pathogenesis of polymorphous light eruption, we studied nine patients with polymorphous light eruption and six healthy persons. Two skin biopsy specimens were obtained from each person, one from previously ultraviolet light-irradiated skin and another one from unirradiated skin. An epidermal cell suspension, skin homogenate, or both were prepared from each specimen. Autologous cultures were made with peripheral blood mononuclear cells combined with irradiated or unirradiated skin homogenate and peripheral blood mononuclear cells combined with irradiated or unirradiated epidermal cell suspension. Cell proliferation was assessed by 3H-thymidine incorporation assay. The response of peripheral blood mononuclear cells to unirradiated epidermal cells or unirradiated skin homogenate was similar in both patients and controls. However, peripheral blood mononuclear cells from patients with polymorphous light eruption showed a significantly increased proliferative response to both irradiated epidermal cells and irradiated skin homogenate. Our results indicate that ultraviolet light increases the stimulatory capability of polymorphous light eruption epidermal cells in a unidirectional mixed culture with autologous peripheral blood mononuclear cells. This suggests that an immune sensitization against autologous ultraviolet light-modified skin antigens occurs in polymorphous light eruption.     

恶性肿瘤相关性皮肌炎/多发性肌炎

皮肌炎/多发性肌炎是一种原因不明的炎性肌肉疾病,常伴发恶性肿瘤,常见的为卵巢癌、乳腺癌、肺癌、胃肠道肿瘤等,而鼻咽癌在亚洲国家较常见.恶性肿瘤相关皮肌炎/多发性肌炎可能的高危因素包括高龄、男性患者、血沉增高、严重的皮损等,其发病机制可能为肿瘤细胞与肌细胞之间相似抗原的交叉免疫反应等.血清自身抗-155抗体是癌症相关性皮肌炎/多发性肌炎的血清学监测指标.     

Terbinafine-induced dermatomyositis: a case report and literature review of drug-induced dermatomyositis

Dermatomyositis, a connective tissue disease syndrome where antibodies to the endothelium of the microvasculature of the skin, muscle and lung are implicated in lesional propagation, is characterized by photodistributed erythema, heliotrope rash, Gottron's papules, muscle weakness and interstitial pulmonary fibrosis. Endotheliotropic viruses and underlying neoplasia are among the inciting triggers. Uncommon drugs, namely the lipid-lowering agents, have been implicated in dermatomyositis. The patient, a 57-year-old man, developed a photodistributed rash and muscle weakness following treatment with the antifungal medication, terbinafine. A skin biopsy was performed, showing an atrophying interface dermatitis with pandermal mucinosis and striking vasculopathic changes including endothelial cell necrosis with denudement and basement membrane zone reduplication. Ultrastructural studies confirmed the presence of endothelial cell injury. Direct immunofluorescent testing showed prominent staining of C5b-9 along the dermal-epidermal junction and within the vasculature. Western blot studies showed strong seroreactivity of his serum to an endothelial-based protein weighing 45,000, a common target described in other microvascular injury-based syndromes. We have shown a temporal association between use of terbinafine and the development of dermatomyositis. The exact basis remains speculative. One potential hypothesis is based on the fact that terbinafine, the active agent in terbinafine, triggers apoptosis of human endothelial cells in culture. Enhanced endothelial cell apoptosis results in the displacement of various cellular antigens creating a state of neoantigenicity; its attendant sequelae is held to be one of anti-endothelial cell antibody formation, a defining pathogenetic event in the evolution of dermatomyositis. The second may be because of the effects of the drug on the promotion of an interferon-rich T-helper-1-dominant cytokine milieu.     

Cutaneous Hyalohyphomycosis Secondary to Paecilomyces Species Treated with Voriconazole in an Immune Compentent Host

Background: Dermatomyositis, a connective tissue disease syndrome where antibodies to the endothelium of the microvasculature of the skin, muscle and lung are implicated in lesional propagation, is characterized by photodistributed erythema, heliotrope rash, Gottron's papules, muscle weakness and interstitial pulmonary fibrosis. Endotheliotropic viruses and underlying neoplasia are among the inciting triggers. Uncommonly drugs, namely the lipid lowering agents, have been implicated in dermatomyositis. Case Report: The patient, a 57‐year‐old male, developed a photodistributed rash and muscle weakness following treatment with the antifungal medication Lamisil. A skin biopsy was performed, showing an atrophying interface dermatitis with pandermal mucinosis and striking vasculopathic changes including endothelial cell necrosis with denudement and basement membrane zone reduplication. Direct immunofluorescent testing showed prominent staining of C5b‐9 along the dermal‐ epidermal junction and within the vasculature. Conclusion: We have shown a temporal association between use of Lamisil and the development of dermatomyositis. Terbinafine, the active agent in Lamisil, promotes apoptosis of human endothelial cells in culture. Enhanced endothelial cell apoptosis results in the displacement of various cellular antigens creating a state of neo‐antigenicity; its attendant sequelae has held to be one of antiendothelial cell antibody formation, a defining pathogenetic event in the evolution of dermatomyositis.     

皮肌炎并发恶性肿瘤患者P53蛋白表达的研究

目的：探讨P53蛋白在皮肌炎并发恶性肿瘤过程中的作用。方法：应用免疫组化技术对皮肌炎患者的肿瘤组织细胞及外周血单个核细胞(PBMCs)P53蛋白的表达进行了检测。结果：1例皮肌炎并发卵巢癌患者的肿瘤细胞及PBMCs均表达P53蛋白，另1例皮肌炎并发鼻咽癌患者的肿瘤细胞、浸润单个核细胞及PBMCs均表达P53蛋白。其它15例未并发恶性肿瘤的皮肌炎患者中有1例PBMCs表达P53蛋白。20例正常对照均阴性。结论：p53基因的种系突变在皮肌炎并发恶性肿瘤中可发能发挥重要作用。     

Studies on Fc-receptor-bearing mononuclear leukocytes from peripheral blood of patients with dermatitis herpetiformis.

Isopaque-Ficoll separated mononuclear cell suspensions from peripheral blood of patients with dermatitis herpetiformis (DH) and healthy controls were investigated by means of a rosette assay for Fc-receptor-bearing leukocytes (EA-RFC), peroxidase staining for monocytes (Pox) and a plaque formation assay (PFC) as well as a 51Cr release assay for cell-mediated cytotoxicity. The cell suspensions were investigated both before and after fractionation on nylon fibre columns. In the patients the mean percentage of PFC in unfractionated cell suspensions was significantly higher than in the controls. In fractionated cell suspensions both the mean percentage of EA-RFC and the mean cytotoxicity index in the 51Cr release assay were significantly lower than in the controls. There were no differences in the percentages of PFC- and Pox-positive cells in fractionated cell suspensions. The results suggest a numerical defect of circulating Fc-receptor-bearing lymphocytes as estimated both by the rosette assay and the 51Cr release assay. These data may reflect a pathogenetic role of these lymphocytes in DH.     

免疫因素在特发性炎症性肌病中的作用机制

特发性炎症性肌炎包括多发性肌炎,皮肌炎及包涵体肌炎,是一组以肌肉组织的炎细胞浸润,肌无力为主要特点,浸润的炎细胞是以T淋巴细胞和巨噬细胞为主,个别也有B淋巴细胞。本文主要综述了T、B淋巴细胞、DC及其它APCs在IIMs中的病理生理作用。     

Circulating natural killer lymphocytes are potential cytotoxic effectors against autologous malignant cells in sezary syndrome patients

Patients with advanced cutaneous T cell lymphoma (CTCL) exhibit profound defects in cell-mediated immunity. Although it has been suggested that Sezary syndrome (SS) patients have a decreased natural killer (NK) lymphocyte activity, nothing has been reported concerning the sensitivity of Sezary cells to NK lymphocyte-mediated cytotoxicity. Peripheral blood NK cells from healthy donors were tested against Sezary tumoral cell lines as well as against freshly isolated Sezary cells. Further, we studied their ability to exhibit antibody -dependent cell-mediated cytotoxicity using either the murine anti-CD158k/KIR3DL2 monoclonal antibody (moAb) AZ158 that specifically recognizes Sezary cells, or the anti-CD52 monoclonal antibody alemtuzumab. The results show that Sezary cell lines are susceptible to NK lymphocyte lysis. More importantly, we found that freshly isolated malignant cells are killed either by IL-2 activated allogeneic NK lymphocytes or when the tumor lymphocyte targets are incubated with an anti-MHC class I F(ab)'2 antibody. Further, anti-KIR3DL2 and anti-CD52 moAb can enhance the NK lysis. Finally, we report that NK lymphocytes isolated from SS patients are potentially cytotoxic lymphocytes against autologous malignant Sezary cells. These findings indicate that antitumor-mediated NK lymphocyte cytotoxic activity can be triggered in patients with CTCL and raise the possibility of developing novel therapeutic strategies by stimulating their innate immunity.     

Impaired responses of peripheral blood mononuclear cells to staphylococcal superantigen in patients with severe atopic dermatitis: a role of T cell apoptosis

Staphylococcus aureus colonization is an almost universal feature of atopic dermatitis. In order to investigate the role of staphylococcal enterotoxin B in the pathogenesis of atopic dermatitis, we assessed the correlation between clinical disease severity and proliferative response of peripheral blood mononuclear cells to staphylococcal enterotoxin B in patients with atopic dermatitis. Peripheral blood mononuclear cells from patients with mild atopic dermatitis showed significantly increased proliferative responses to staphylococcal enterotoxin B compared to controls. In contrast, peripheral blood mononuclear cells from patients with severe atopic dermatitis showed markedly suppressed proliferative responses. Additionally, longitudinal evaluation of peripheral blood mononuclear cell samples from the same patient demonstrated that proliferative responses were suppressed only at times of severe disease exacerbation. Mixing experiments, using autologous T cells and antigen presenting cells that were isolated at different time points from the same patient, demonstrated that T cells of severe atopic dermatitis patients were dysfunctional, but their antigen presenting cell function remained intact. We found no significant differences of interleukin-2 levels in the culture supernatants between healthy controls and atopic dermatitis groups. Fluorescence-activated cell sorter analysis for APO2.7 antigen, an early apoptosis cell marker, demonstrated that approximately 60% of staphylococcal-enterotoxin-B-stimulated T cells expressed APO2.7 antigen in severe atopic dermatitis cases. By contrast, 5%-20% of T cells expressed APO2.7 after staphylococcal enterotoxin B stimulation in cases of mild atopic dermatitis and in healthy controls. Nuclear staining with Hoechst 33258 also showed approximately 40% apoptotic cells in the CD19-CD16-PBMC of severe atopic dermatitis patients, compared with only 5%-10% in the mild atopic dermatitis group and in healthy controls. Blocking monoclonal antibody to Fas ligand partially prevented the staphylococcal-enterotoxin-B-induced apoptosis detected by APO2.7 expression and Hoechst 33258 staining. Suppressed proliferation of peripheral blood mononuclear cells in severe atopic dermatitis patients may be secondary to T cell death by apoptosis. These results suggest that an infection of S. aureus producing staphylococcal enterotoxin B may play a role in aggravation of atopic dermatitis by inducing apoptosis in T cells.     

Cutaneous changes of dermatomyositis precede muscle weakness

A retrospective review of 50 patients with dermatomyositis was performed to determine the temporal relationship between onset of muscle weakness and skin involvement. We found that cutaneous changes sometimes preceded muscle weakness more than a year before the onset of muscle weakness. These findings suggest that the characteristic dermatomyositis eruption without muscle weakness should not preclude a diagnosis of dermatomyositis, and these cases should be carefully followed.     

无肌病性皮肌炎一例

患者,女,21岁。面部、躯干、四肢皮肤红斑、丘疹伴脱发6年,皮损基本损害为红斑,双眼睑见明显水肿性紫红斑,其他部位皮损为弥漫性对称性暗紫红色斑,双手可见Gottron丘疹。肌力正常。皮损组织病理检查：表皮轻度角化过度,基底膜增厚,真皮浅层水肿,血管周围单一核细胞浸润。诊断：无肌病性皮肌炎。给予泼尼松联合甲氨蝶呤治疗2个月后皮损明显好转。     

Blood lymphocyte subpopulations in Mycosis fungoides and their functions in vitro

T-lymphocyte subpopulations, PWM stimulated in vitro Ig production by mononuclear cells, and spontaneous cell-mediated cytotoxicity (SCMC) were determined in patients with Mycosis fungoides (MF) as well as in sex- and age-matched normal controls. We were able to confirm our earlier findings of a significantly higher frequency of T-lymphocytes with Fc-receptors for IgG (Tg cells) and a proportion of T lymphocytes with Fe receptors for IgM (Tm cells) not significantly differing from normal controls. As defined by monoclonal antibodies, the proportion of T-helper cells (Leu3A) was significantly lower, whereas the T-suppressors cells (Leu2A) were in the range of normal controls. The PWM stimulated and spontaneous Ig synthesis in vitro was lower in MF patients whereas there was no difference in the SCMC activity, in comparison with normal controls. The lower frequency of T-helper cells in the peripheral blood may be explained by their migration to the skin.