Decongestion improves cell-mediated immunity in postmastectomy arm lymphoedema: a pilot study.

BACKGROUND: Chronic lymphoedematous limbs have an increased propensity for infections and primary or secondary malignant tumours. It has been attributed to suppressed delayed-type hypersensitivity measured in lymphoedemas related to Stewart-Treves syndrome, Kaposi's sarcoma or breast cancer treatment. Cell-mediated immunity is an effective defence mechanism against bacteria, fungi, viruses and tumour cells. OBJECTIVE: We aimed to examine whether decongestive lymphoedema therapy could improve cell-mediated immunity in breast cancer treatment-related lymphoedema (BCRL). METHODS: Eight women with unilateral BCRL were included in this study. At baseline, tuberculin skin test (TST) was performed on the volar surfaces of the forearms of the affected and non-affected sides using 0.5, 1 and 5 tuberculin units in the form of three consecutive injections with 3-cm spaces in-between, and arm volumes were measured using the Kuhnke's disc model. Decongestive lymphatic therapy was given to swollen arms in 10 consecutive working days. At the end of intensive decongestion, TST on affected side and bilateral volumetry were repeated. RESULTS: Baseline test using undiluted (5 units) and fivefold diluted (1 unit) tuberculin solutions has shown significant differences (P < 0.05) between the mean sizes (11.81 ± 2.32 and 7.75 ± 1.92; 7.12 ± 1.12 and 5.12 ± 0.91 respectively) in favour to healthy arms. Post therapeutically, the mean sizes were significantly increased (P < 0.05) in the dilutions of 1 : 1 and 1 : 5 (7.75 ± 1.92 and 10.56 ± 1.23 mm, 5.12 ± 0.91 and 5.93 ± 1.74 mm respectively). CONCLUSION: Significant increase in TST sizes suggests that decongestive lymphatic therapy is able to partially restore impaired cellular immune function in BCRL.

Intermittent pneumatic compression acts synergistically with manual lymphatic drainage in complex decongestive physiotherapy for breast cancer treatment-related lymphedema.

The application of intermittent pneumatic compression (IPC) as a part of complex decongestive physiotherapy (CDP) remains controversial. The aim of this study was to investigate whether the combination of IPC with manual lymph drainage (MLD) could improve CDP treatment outcomes in women with secondary lymphedema after breast cancer treatment. A randomized study was undertaken with 13 subjects receiving MLD (60 min) and 14 receiving MLD (30 min) plus IPC (30 min) followed by standardized components of CDP including multilayered compression bandaging, physical exercise, and skin care 10 times in a 2-week-period. Efficacy of treatment was evaluated by limb volume reduction and a subjective symptom questionnaire at end of the treatment, and one and two months after beginning treatment. The two groups had similar demographic and clinical characteristics. Mean reductions in limb volumes for each group at the end of therapy, and at one and two months were 7.93% and 3.06%, 9.02% and 2.9%, and 9.62% and 3.6%, respectively (p < 0.05 from baseline for each group and also between groups at each measurement). Although a significant decrease in the subjective symptom survey was found for both groups compared to baseline, no significant difference between the groups was found at any time point. The application of IPC with MLD provides a synergistic enhancement of the effect of CDP in arm volume reduction.

The prevalence of melanocytic naevi among schoolchildren in South Hungary.

BACKGROUND: Malignant melanoma is an increasing public health problem worldwide; accordingly, identification of the constitutional and environmental factors which contribute to the development of the disease, and hence identification of the individuals at high risk of melanoma, is an indispensable step in all primary prevention efforts. OBJECTIVES: This paper aims to assess the prevalence of different pigmented lesions among schoolchildren and to investigate their relationship with phenotypic pigmentary characteristics, sun exposure and other factors. PATIENTS/METHODS: A cross-sectional study was performed in two secondary schools in Szeged, Hungary. A total of 1320 schoolchildren, aged 14 to 18 years, underwent a whole-body skin examination. A standardized questionnaire was used to collect data on phenotypic, sun exposure and other variables. RESULTS: One to 10 common melanocytic naevi were found in 27% of the participants, and the naevus numbers were in the range of 10-100 in 67%; 5.4% of them had more than 100 common melanocytic naevi. The prevalence of clinically atypical naevi was 24.3%. Statistically significant associations were found between the number of pigmented lesions and gender, hair colour, eye colour, skin phototype, a history of severe painful sunburns and a family history of a large number of melanocytic naevi. CONCLUSION: Our study population displayed a markedly high prevalence of clinically atypical melanocytic naevi. Moreover, a considerable proportion of the investigated individuals had multiple common melanocytic naevi. Since the presence of a large number of melanocytic naevi is a strong predictor for future melanoma development, health educational programmes on melanoma prevention should be aimed at young age groups.

Adjunctive role of manual lymph drainage in the healing of venous ulcers: A comparative pilot study.

Compression therapy plays a pivotal role in the treatment of venous leg ulcers and clinical observations include lymph stasis as contributing to the maintenance of chronic wounds. This finding raises the question whether further improvement in lymph circulation with manual lymph drainage (MLD) as a part of complex decongestive physiotherapy (CDP) can improve ulcer healing. We examined whether CDP improves healing of venous leg ulcers and compared the efficacy of CDP with that of multilayered compression with short-stretch bandages. Eight patients (mean age: 64.8 years, mean ulcer area: 23.07 cm2, duration of ulcers: 25.37 months) were treated with a 5-day-course of CDP and 9 patients (mean age: 70.77 years, mean ulcer area: 21.47 cm2, duration of ulcers: 15.8 months) were included in a 10-day-course of CDP. Control goup consisted of 9 patients (mean age: 56.33 years, mean ulcer area: 13.87 cm2, duration of ulcers: 6.11 months) receiving multilayered compression. Wound surface measurement was carried out on days 5 and 10 and ulcer area reduction rate was calculated as area (initial)-area (final)/time unit. There was no statistical difference between the 5-daycourse of CDP and compression of the same duration regarding ulcer healing (t=-1.62, df=15, p= 0.125). A 10-day-course of CDP significantly increased ulcer healing compared to compression of the same duration (t=-2.42, df=16, p= 0.039). Our preliminary results suggest that MLD as a part of CDP supports healing of venous leg ulcers.

Platelet-activating factor antagonists (BN 52021 and BN 50730) inhibit tumor necrosis factor-alfa-mediated cytotoxicity on murine L929 tumor cells.

Tumor necrosis factor (TNF)-alfa has been described as a mononuclear phagocyte-produced cytotoxin that causes the necrosis and regression of some tumors. The mechanism of the cytotoxicity and the basis for the differential cytotoxic effects of TNF against cells of various origin remains unclear. It has also been reported, that murine TNF stimulates the production of platelet-activating factor (PAF) by cultured peritoneal macrophages, and that PAF enhances TNF production by alveolar macrophages. Furthermore, it is known that the synthesis and release of PAF are inhibited by plasma proteinase inhibitors. This study was devoted to investigate the effects of two specific PAF antagonists (BN 52021 and 50730), and a proteinase inhibitor (aprotinin; GordoxR) on the TNF-induced cytotoxicity in L929 murine fibroblasts. Our present findings indicate that TNF-induced cytotoxicity is inhibited in a dose-dependent manner by the PAF antagonists studied and by the kallikrein inhibitor aprotinin. These findings provide further evidence suggesting that PAF might be involved in the process of the TNF-alfa-induced cytotoxicity of L929 mouse fibroblasts.

A comparative study of the shedding process mouse erythrocyte binding receptors on peripheral blood lymphocytes from healthy donors and patients suffering from chronic lymphocytic leukemia.

The shedding process of the mouse erythrocyte binding receptors of human peripheral lymphocytes of a healthy donor group was compared to that of patients with chronic lymphocytic leukemia. The shedding process exhibited significant differences with respect to the groups tested, suggesting that the mouse erythrocyte binding receptors were altered during leukemic transformation or that the leukemic cells were fixed in a stage of maturation.

Suppressor function of peripheral blood mononuclear cells in patients with psoriasis vulgaris.

The suppressor activity of peripheral blood mononuclear cells was investigated in 25 patients with psoriasis vulgaris. In the psoriatic patients the suppressor activity was found to be significantly lower than in the control group, which suggests that the weak suppressor activity may play a role in the pathogenesis of this disease.

A mannose-binding receptor is expressed on human keratinocytes and mediates killing of Candida albicans.

Human keratinocytes are known to kill Candida albicans in vitro, but the mechanism of killing is not yet understood. Here, we demonstrate that spontaneous, ultraviolet-B-light-induced, alpha-melanocyte-stimulating-hormone-induced, and interleukin-8-induced Candida killing by keratinocytes can be inhibited with mannan and mannosylated bovine serum albumin (Man-BSA). A polyclonal goat serum raised against the human macrophage mannose receptor stained suprabasal keratinocytes, but no staining was observed on keratinocytes with a monoclonal antibody (mAb15) specific for the human macrophage mannose receptor. Mannose-affinity chromatography of keratinocyte extract isolated a 200 kDa protein, and on the Western blot the goat antiserum reacted with a 200 kDa protein. In radioligand binding studies, the binding of 125I-Man-BSA to human keratinocytes was inhibited by mannan in a concentration-dependent manner. Analysis of the binding revealed a single class keratinocyte mannose receptor with a KD of 1.4 x 10(-8) M and a Bmax of 1 x 10(4) binding sites per cell. The binding of 125I-Man- BSA to keratinocytes proved to be time-dependent, acid-precipitable, and Ca2+- and trypsin-sensitive. After trypsinization the receptors underwent a rapid recovery at 37 degrees C. These results demonstrate the presence of mannose receptor on human keratinocytes, and its active involvement in the killing of Candida albicans.

Histidine decarboxylase expression in human melanoma.

Histamine has been implicated as one of the mediators involved in regulation of proliferation in both normal and neoplastic tissues. Histidine decarboxylase, the only enzyme that catalyzes the formation of histamine from L-histidine, is an essential regulator of histamine levels. In this study, we investigated the gene and protein expression of histidine decarboxylase in melanoma. Reverse transcriptase polymerase chain reaction and in situ hybridization studies of WM-35, WM-983/B, HT-168, and M1 human melanoma cell lines both resulted in positive signals for histidine decarboxylase messenger RNA. A polyclonal chicken antibody was developed against human histidine decarboxylase and protein expression was confirmed by western blot analysis of the cell lysates, revealing a predominant immunoreactive band at approximately 54 kDa corresponding to monomeric histidine decarboxylase. Protein expression of histidine decarboxylase was also shown by flow cytometric analysis and strong punctate cytoplasmic staining of melanoma cell lines. Moreover, both primary and metastatic human melanoma tissues were brightly stained for histidine decarboxylase. When compared with the very weak or no reactions on cultivated human melanocytes both western blot and immunohistochemical studies showed much stronger histidine decarboxylase expression in melanoma cells. These findings suggest that expression of histidine decarboxylase is elevated in human melanoma.

Immune-associated surface markers of human keratinocytes.

The results of a number of investigations have proved that human keratinocytes (HKs) possess the ability to synthesize and express cell surface moieties characteristic of effector and/or accessory cells of the immune system. The present paper summarizes the known immune cell surface features of HKs, reflecting their stage of activation and differentiation. The surface and functional characteristics of HKs suggest their monocyte/macrophage behavior, which fits in well with the presumed active involvement of HKs in the skin immune system.

Identification of a soluble interleukin-8 inhibitor in the supernatant of polymorphonuclear leukocytes.

Interleukin-8 (IL-8) plays a crucial role in the pathogenesis of inflammatory and hyperproliferative diseases in various organs. The purpose of the present investigation was to establish whether there is any naturally occurring inhibitor of IL-8. Here we demonstrate that an IL-8 inhibitor (IL-8INH) is present in the supernatant of polymorphonuclear (PMN) leukocytes. The release of IL-8INH could be increased by stimulating the PMN leukocytes by concanavalin A. IL-81NH blocks the IL-8-induced chemotaxis and Candida albicans killing activity of PMN leukocytes and epidermal cells in vitro, and IL-8-induced neutrophil infiltration in the mouse ear in vivo. The mechanism of action of IL-8INH involves blocking of 125I-IL-8 binding to the IL-8 receptor. Binding of 125I-IL-8 to neutrophils could not be displaced by the IL-8INH, however, preincubation of 125I-IL-8 with IL-8INH increased binding inhibition, suggesting an interaction between IL-8 and the inhibitor. Crosslinking of 125I-IL-8 to IL-8INH shows that IL-8INH binds specifically to 125I-IL-8, and the IL-8INH protein has an apparent molecular weight of 52 kDa in sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The partial purification of the IL-8INH on DEAE-Sephadex anion-exchange chromatography column also suggests a 50-60-kDa inhibitor protein which blocks IL-8-induced effects on neutrophils by binding to IL-8.

The interleukin-8 receptor: a potential target for antipsoriatic therapy?

Interleukin-8 is assumed to play a central role in the pathogenesis of psoriasis. Since an increased expression of the interleukin-8 receptor has been observed both in polymorphonuclear leukocytes and in affected psoriatic epidermis, we were interested in whether the interleukin-8 receptor could be a molecular target of antipsoriatic compounds. Cyclosporine, calcitriol, calcipotriol or dithranol caused a dose-dependent decrease in interleukin-8 binding to cultured human keratinocytes, while interleukin-8 binding to granulocytes was not affected. In addition, the interleukin-8-induced human leukocyte antigen-DR (HLA-DR) expression of keratinocytes was nearly completely blocked by treatment of the cells with these substances. The inhibition of the keratinocyte interleukin-8 receptor and its function by antipsoriatic drugs may contribute to their therapeutic action.

Topical tacrolimus is not effective in chronic plaque psoriasis. A pilot study.

BACKGROUND: Cyclosporine for the treatment of psoriasis constitutes a new approach. Alternative systemic cyclosporine derivatives have been studied to find an immunosuppressive drug with fewer adverse effects. Tacrolimus is one of these new immunosuppressive drugs. Systematically, it has been proven effective in treating psoriasis. A topical formulation of tacrolimus is attractive because it has fewer adverse effects and is useful for a large group of patients. We report for the first time on the efficacy of nonocclusive topical tacrolimus in the treatment of psoriasis. OBSERVATIONS: After a washout phase of 2 weeks, patients were randomized to receive 0.005% calcipotriol ointment twice daily, placebo ointment once daily, or 0.3% tacrolimus ointment once daily. One psoriatic plaque was treated with a surface area of 40 to 200 cm2. Efficacy was estimated using the local psoriasis severity index. The reduction in the local psoriasis severity index score after 6 weeks was 62.5% in the calcipotriol group, 33.3% in the tacrolimus group, and 42.9% in the placebo group. CONCLUSIONS: There was no statistically significant difference between the efficacy of tacrolimus and placebo ointment (P = .77). Calcipotriol ointment, applied twice daily, had a better effect than tacrolimus ointment and placebo ointment once daily.

Enhancement of mouse erythrocyte rosette formation of human lymphocytes by interferon gamma.

Interferon-gamma enhances the mouse erythrocyte rosette formation of human peripheral blood mononuclear cells in a dose-dependent way. Pretreatment of the cells with colchicin abolishes the enhancing effect. An increased expression of the mouse erythrocyte binding receptor may underlie the phenomenon.

Pharmacological studies on dithranol-induced irritative dermatitis in mice.

The effects of different pharmacological substances on dithranol-induced irritative dermatitis were studied in mice. Pretreatment of the animals with a specific platelet activating factor (PAF) antagonist, BN 52021, significantly reduced the ear swelling in a dose-dependent manner. The cyclooxygenase inhibitor indomethacin, the antihistamine clemastine, and the anti-oxidant superoxide dismutase also proved to be effective in the reduction of the dermatitis. The results provide evidence of the coinvolvement of PAF, prostaglandins, histamine, and reactive oxygen radicals in dithranol-induced irritative dermatitis in mice.

Study of phagocytic function with a quantitative nitroblue-Tetrazolium (NBT) reduction test in diabetes mellitus.

In 40 patients suffering from diabetes mellitus and in 18 healthy volunteers the phagocytic function was investigated using a quantitative NBT test. The NBT reduction was significantly lower in diabetes than in healthy donors irrespective of affliction with other dermatoses. A significant correlation was found between the severity and hence the insulin dependency of diabetics and the NBT-reductive capacity.

Evidence for cell-mediated autoimmunity in patients with pemphigus vulgaris and bullous pemphigoid.

Lymphoid cells from 4 of 5 patients diagnosed as pemphigus vulgaris (PV) and 6 of 7 patients diagnosed as bullous pemphigoid (BP) demonstrated specific cell-mediated immunity by the production of migration inhibitory factor (MIF) in the presence of autologous epidermal saline extracts. Clinical treatment of these patients with immunosuppressive agents resulted in a state of unresponsiveness of their lymphoid cells to similar concentrations of the antigen. Controls consisted of lymphoid cells from patients with bullous burns or various drug allergies which failed to show significant MIF production in the presence of autologous skin extract. These studies suggest that both PV and BP patients posses cell-mediated immunity (CMI) to their own autologous tissue antigens and this CMI may play a role in the pathogenesis of these diseases.

Polymorphonuclear granulocyte chemotaxis and chemotactic factor generation by concanavalin A-stimulated peripheral blood mononuclear cells in patients with psoriasis.

Polymorphonuclear leukocytes from patients with psoriasis demonstrated a significantly enhanced chemotactic responsiveness to zymosan-activated human serum as compared to granulocytes from healthy volunteers. Furthermore, psoriatic peripheral blood mononuclear cells stimulated with concanavalin a produced an increased amount of lymphocyte derived chemotactic factor (LDCF) as compared to that in the case of healthy persons. The LDCF proved to be chemokinetic for the psoriatic granulocyte. It is postulated that these two phenomena may play a role in the pathogenesis of psoriasis.

Serum factors regulate the expression of the proliferation-related genes alpha5 integrin and keratin 1, but not keratin 10, in HaCaT keratinocytes.

In the highly coordinated programme of gene expression during keratinocyte proliferation and differentiation, alpha5 integrin and keratins 1 and 10 (K1/K10) may play important regulatory roles. We were interested in seeing whether, in continuously growing, immortalized HaCaT keratinocytes, similar to normal keratinocytes, the expression of alpha5 integrin and K1/K10 was related to cell proliferation and differentiation. After release from cell quiescence the expression of alpha5 integrin, both at the mRNA and protein levels, was upregulated in the cells. At the same time, K1/K10 mRNA and protein expression decreased dramatically, while the mRNA for D1 cyclin became detectable, and the cells became highly proliferative. These findings indicate that alpha5 integrin and K1/K10 are involved in the regulation of HaCaT proliferation and differentiation, as in normal keratinocytes. However, HaCaT cells are different from normal keratinocytes in their ability to lose K1/K10 expression. There is no evidence that the expression of K1/K10 can be reversed in normal keratinocytes. This ability of dedifferentiation might be a unique feature of HaCaT cells and may be a key component of their immortalized nature. We also found that serum factors regulate mRNA expression of alpha5 integrin and K1, but not of K10, in HaCaT cells. This information could be relevant to the understanding of normal epidermal differentiation.