Biopsy Specimens Of Normal Skin Research Articles

Expression of JAK/STAT signalling in bullous pemphigoid.

In the present study, we characterise the expression of JAK/STAT signalling in Bullous Pemphigoid (BP). Punch biopsy specimens of normal skin, lesional and perilesional skin of patients diagnosed with biopsy proven BP were utilised for immunoperoxidase labelling with commercially available antibodies for Phosphorylated STAT 1,2,3,5A and 6 (Genesearch, USA). Quantitative analysis demonstrated that in comparison to normal skin, phosphorylated STAT 2, 3, 5A and 6 were overexpressed in BP.

Comparison of the tissue expressions of glutathione S transferase isoenzymes among patients with morphea and healthy controls: A preliminary study.

Morphea is an inflammatory connective tissue disorder, which is characterized by sclerosis in skin and subcutaneous tissues with a chronic progress. The oxidative stress in pathogenesis of sclerosing diseases was proposed in several studies with conflicting results. To explore the tissue expressions of Glutathione S transferase (GST) isoenzymes in patients with morphea and compare these expressions with healthy controls. Twenty-two morphea patients and 20 sex and age matched healthy controls were enrolled in this study. Four millimeter punch biopsies were performed from the active sclerotic plaques of morphea patients. Tissue samples of control group were obtained from nonlesional normal skin biopsy specimens. The protein expressions of GST isoenzymes were analyzed immunohistochemically. Tissue expressions of GSTP1, GSTT1, and GSTA1 isoenzymes in morphea patients were found to be significantly higher than in control tissues. There was no significant difference in GSTM1 isoenzyme expression between the two groups. The increased tissue expressions of GSTA1, GSTP1, and GSTT1 isoenzymes in morphea may represent the activated GST enzymes in response to excessive free radical formation and may also support the hypothesis of increased oxidative stress in morphea etiopathogenesis.

Effects of Combined Treatment With Arsenic Trioxide and Itraconazole in Patients With Refractory Metastatic Basal Cell Carcinoma.

Tumor resistance is an emerging problem for Smoothened (SMO) inhibitor-treated metastatic basal cell carcinoma (BCC). Arsenic trioxide and itraconazole antagonize the hedgehog (HH) pathway at sites distinct from those treated by SMO inhibitors. To determine whether administration of intravenous arsenic trioxide and oral itraconazole in patients with metastatic BCC is associated with a reduction in GLI1 messenger RNA expression in tumor and/or normal skin biopsy samples. Five men with metastatic BCC who experienced relapse after SMO inhibitor treatment underwent intravenous arsenic trioxide treatment for 5 days, every 28 days, and oral itraconazole treatment on days 6 to 28. Data were collected from April 10 to November 14, 2013. Follow-up was completed on October 3, 2015, and data were analyzed from June 5 to October 6, 2015. The primary outcome was the change in messenger RNA levels of the GLI family zinc finger 1 (GLI1) gene (HH-pathway target gene) in biopsy specimens of normal skin or BCC before and after treatment. Secondary objectives were evaluation of tumor response and tolerability. Of the 5 patients (mean [SD] age, 52 [9] years; age range, 43-62 years), 3 completed 3 cycles of treatment and 2 discontinued treatment early owing to disease progression or adverse events. Adverse effects included grade 2 transaminitis and grade 4 leukopenia with a grade 3 infection. Overall, arsenic trioxide and itraconazole reduced GLI1 messenger RNA levels by 75% from baseline (P < .001). The best overall response after 3 treatment cycles was stable disease in 3 patients. Targeting the HH pathway with sequential arsenic trioxide and itraconazole treatment is a feasible treatment for metastatic BCC. Although some patients experienced stable disease for 3 months, none had tumor shrinkage, which may be owing to transient GLI1 suppression with sequential dosing. Continuous dosing may be required to fully inhibit the HH pathway and achieve clinical response.

Evaluation of heparanase and matrix metalloproteinase‐9 in patients with cutaneous malignant melanoma

Elevated heparanase and matrix metalloproteinase (MMP)-9, frequently found in human cancer, is a major cause of degradation of the extracellular matrix (ECM) and basement membrane (BM), thus facilitating tumor cell migration and invasion. Although a lot of work has been done, the role of heparanase and MMP-9 has not been delineated in skin cancer progression. The purpose of this study was to do such an exploration. To investigate the role of heparanase and MMP-9 in cutaneous malignant melanoma (CMM) development, we performed immunohistochemical analysis to detect the alternation of these two factors in paraffin-embedded biopsy specimens of normal skin, junctional nevi and CMM. It is interesting to note that the expression profile of heparanase and MMP-9 was similar. Contrary to negative staining in normal skin, overexpression of heparanase and cytoplasmic MMP-9 was observed in as many as 70% of CMM, whereas only 10% of the junctional nevi exhibited faint staining (P = 0.0005, P = 0.0000). Considering the lymph node (LN) metastasis, the expression of the two factors is significantly higher in LN-positive lesions than that in LN-negative lesions (P = 0.0295, P = 0.0013). Meanwhile, there was positive correlation between the expression of MMP-9 and heparanase (r = 0.689, P = 0.003). The first expression of MMP-9 and heparanase occurs at benign lesions. However, the significantly increased expression in advanced CMM stages, particularly in LN-positive metastasis lesions, might synergistically contribute to degradation of ECM and BM, therefore promoting carcinogenesis and metastasis.

Expression of P27, Ki67 and P53 in Squamous Cell Carcinoma, Actinic Keratosis and Bowen Disease

This study aims at evaluating the expression of P27, Ki67 and P53 in Squamous Cell Carcinoma (SCC), Actinic Keratosis (AK) and Bowen Disease (BD) specimens. In an analytic-descriptive setting, skin biopsy specimens of 45 patients were evaluated in three 15-case groups including BD, AK and SCC specimens. Fifteen normal skin biopsy specimens were obtained and used as the control group. Immunohistochemical staining was performed in all the specimens and the expression rates and patterns of Ki67, P27 and P53 were determined. The results were compared between the four groups. Ki67 was expressed in 0.8, 23.7, 12.3 and 19.3% of the cells in the normal skin, AK, BD and SCC groups, respectively. No significant difference was seen between the three pathological conditions regarding the expression rate of Ki67. P27 was positive in 23.4, 26.2, 25.9 and 4.5% of specimens in the normal skin, AK, BD and SCC groups, respectively. This rate was significantly the lowest in the SCC group. P53 expression was detected in 26.6, 41.8 and 54.6% of the assessed cells in the AK, BD and SCC groups, respectively. There was no expression of P53 in the normal skin specimens. This rate was significantly the highest again in the SCC group. Based on these results, the quantitative and qualitative (pattern of distribution) evaluation of the expressions of Ki67, P27 and P53 may be helpful in differentiating malignant and premalignant epidermal lesions, particularly in unsatisfactory or fragmented specimens.

Association between enhanced type I collagen expression and epigenetic repression of the FLI1 gene in scleroderma fibroblasts

Scleroderma (systemic sclerosis; SSc) is an autoimmune disease characterized by vasculopathy and widespread organ fibrosis. Altered fibroblast function, both in vivo and in vitro, is well documented and illustrated by augmented synthesis and deposition of extracellular matrix proteins. We undertook this study to investigate the possibility that epigenetic mechanisms mediate the emergence and persistence of the altered SSc fibroblast phenotype. The effects of DNA methyltransferase and histone deacetylase inhibitors on collagen expression and the level of epigenetic mediators in fibroblasts were examined. The effects of transient transfection of SSc fibroblasts with FLI1 gene and normal cells with FLI1 antisense construct on collagen expression were determined. The methylation status of the FLI1 promoter was tested in cultured cells and in SSc and normal skin biopsy specimens. Increased levels of epigenetic mediators in SSc fibroblasts were noted. The addition of epigenetic inhibitors to cell cultures normalized collagen expression in SSc fibroblasts. The augmented collagen synthesis by SSc fibroblasts was linked to epigenetic repression of the collagen suppressor gene FLI1. Heavy methylation of the CpG islands in the FLI1 promoter region was demonstrated in SSc fibroblasts and skin biopsy specimens. The results of this study indicate that epigenetic mechanisms may mediate the fibrotic manifestations of SSc. The signal transduction leading to the SSc fibrotic phenotype appears to converge on DNA methylation and histone deacetylation at the FLI1 gene.

Differential expression of cornified cell envelope precursors in normal skin, intraorally transplanted skin and normal oral mucosa.

Skin flaps have routinely been used as substitutes for oral mucosa after extensive resection of oral tissues. However, it remains unknown how the transplanted skin flaps perform as a host defence in the new environment of the oral cavity. To evaluate the expression of cornified cell envelope (CCE) precursors in pretransplanted (normal) skin, intraorally transplanted skin and normal oral mucosa, because CCEs are highly responsible for a protective barrier in each type of epithelium. We used immunohistochemistry and immunoelectron microscopy to examine the expression of CCE precursors, small proline-rich protein (SPR) 2 and 3 and loricrin, in biopsy specimens of normal skin, transplanted skin and normal oral mucosa, including buccal and lingual (non-keratinized) mucosae, and palatal (keratinized) mucosa. Transplanted skin flaps were classified into two groups. About two-thirds of the transplanted skin flaps displayed a reddish appearance and were devoid of the stratum corneum (SC) together with a psoriasiform inflammatory tissue reaction. Others showed a native appearance, retaining the SC. While SPR2 expression was limited to the stratum granulosum (SG) in both normal and transplanted skin retaining the SC, it extended to the stratum spinosum (SS) of the transplanted skin lacking the SC and that of the normal oral mucosa. Although SPR3 expression was not found in normal skin or in the transplanted skin retaining the SC, it was strongly expressed in the SS of the transplanted skin lacking the SC and the non-keratinized oral mucosa, and in the SS and SG of the keratinized oral mucosa. Loricrin, which was expressed in the SG of normal skin, the transplanted skin retaining the SC and the keratinized oral mucosa, was not detected in the transplanted skin lacking the SC or in the non-keratinized oral mucosa. Immunoelectron microscopy confirmed the ultrastructural localization of SPR3 directly under the cytoplasmic membrane of keratinocytes of the transplanted skin lacking the SC and that of the oral mucosa. The altered expression of SPR2, SPR3 and loricrin reflects the possible adaptation of epidermal keratinocytes in the new environment of the oral cavity.

Bednár tumor associated with dermal melanocytosis: melanocytic colonization or neuroectodermal multidirectional differentiation?

Neuroectodermal differentiation or melanocytic colonization are the opposing theories of histogenesis for the Bednár tumor or pigmented dermatofibrosarcoma protuberans (DFSP). A 31-year-old African-American woman presented with a 2-cm blue-black shoulder nodule of 1-year duration. Punch biopsy revealed a CD34+, Factor XIIIa-DFSP, harboring numerous, pigmented spindle S100+, Mart-1+ and HMB-45+ cells. Subsequent wide excision demonstrated pigmented dendritic and spindled cells widely scattered throughout the dermis of the 3-cm excisional margins and punch biopsy specimens of normal skin from both shoulders. This latter process was interpreted as dermal melanocytosis (nevus of Ito). The dermal pigmented spindle cells were Mart-1+ and CD34-, and were associated with non-pigmented CD34+, cytologically banal spindle cells, which were more numerous in the excisional margins than the contralateral shoulder. Reported herein is a singular case of Bednár tumor associated with dermal melanocytosis. Although the coexistence of these processes implicates colonization of the DFSP by constituent dermal melanocytes, the mixed immunophenotype (CD34+ or Mart-1+ cells) of dispersed dermal spindle cells hints at the possibility of a common cell of origin: the putative neuromesenchymal cell. In effect, the Bednár tumor could represent one part of a spectrum of neural crest-derived dermal tumors that includes dermal melanocytosis, cellular blue nevus and conventional DFSP.

Morphometric analysis of ultrastructural vascular changes in CADASIL: analysis of 50 skin biopsy specimens and pathogenic implications.

Cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) is a systemic vascular disease caused by Notch 3 gene mutations. On electron microscopy a specific granular osmiophilic material (GOM) is found surrounding the vascular smooth muscle cells. In 1993, we first proposed the use of skin biopsy to diagnose patients and to identify relatives of patients with CADASIL. We analyze here our experience with skin biopsies from 50 patients with CADASIL and compare the findings with those of 20 normal skin biopsy specimens. A morphometric analysis of skin vessel morphology on electron microscopy was performed by systematic measurements of several blood vessel diameters, as well as of areas of lumen, endothelial cell and smooth muscle cell cross-sectional areas, vessel wall area, arterial media and extracellular matrix areas. We found relative absence of stenosis but marked destruction of smooth muscle cells, resulting in decrease of vessel wall thickness and loss of extracellular matrix area, producing vessel wall weakness. Similar changes were also observed in brain arterioles from 5 patients with CADASIL. Our results suggest that hypotonicity of the arteriolar tree may constitute an important pathogenetic mechanism in CADASIL. Other than hypotonicity, the early and severe destruction of smooth muscle cells may potentially result in decreased secretion of vascular endothelial growth factor, loss of vascular permeability and damaging hemodynamic consequences. Blood vessel morphology of skin vessels correlated well with changes in brain arterioles. Vascular morphology in skin biopsy samples contributes to our understanding of the pathogenesis of CADASIL. It could be important to perform skin biopsies in future therapeutic trials of CADASIL as a direct measure of therapeutic effectiveness.

Detection of type 1 cytokines in discoid lupus erythematosus.

Although multiple studies suggest a dysregulated T-cell cytokine production in systemic lupus erythematosus, the cytokine profile in discoid lupus erythematosus (DLE) lesions is unknown. To characterize the cytokine profile in DLE by immunohistochemical and molecular methods, and to investigate the role of cytokines in the pathogenesis of DLE. Patients were evaluated clinically, and biopsy specimens of lesional skin were examined by light microscopy. Reverse transcriptase-polymerase chain reaction and immunohistochemical analysis were performed on 11 biopsy specimens. We investigated the presence of interleukin (IL) 2, interferon gamma (IFN-gamma), IL-4, tumor necrosis factor alpha, (TNF-alpha), and IL-1beta messenger RNA (mRNA) in 8 biopsy specimens of DLE and compared it with 3 biopsy specimens of normal skin. Academic referral research hospital. Eight consecutive patients with a clinical and histologic diagnosis of DLE. Localized DLE was found in 7 patients and widespread in 1. During the 4 years of the investigation, none of the patients developed systemic lupus erythematosus. We found significantly elevated levels of IL-2 and IFN-gamma mRNA in all 8 biopsy specimens of DLE; in contrast, no transcripts of IL-2 or IFN-gamma were detected in 3 biopsy specimens of normal skin (P<.01). Similarly, elevated levels of TNF-alpha mRNA were detected in 8 DLE biopsy specimens, while no TNF-alpha mRNA was detected in 3 biopsy specimens of normal skin (P<.01). No IL-4 or IL-1 beta mRNA was detected in 8 biopsy specimens of DLE lesional skin and 3 biopsy specimens of normal patient skin. Immunohistochemical analysis showed increased staining for IL-2 and IFN-gamma receptors, while no detectable IL-4 receptor was found. No cytokine mRNA or cytokine receptor protein was detected in biopsy specimens of normal skin. These findings suggest that DLE is associated with type 1 cytokines characterized by the expression of IL-2 and IFN-gamma. Type 1 cytokines may be critical for induction, development, and maintenance of DLE.

Expression of E-cadherin, [formula omitted], and CD44V 6 and the subcellular localization of E-cadherin and CD44V 6 in normal epidermis and basal cell carcinoma

Basal cell carcinoma (BCC) of the skin is a locally invasive, rarely metastasizing epithelial tumor. In the current study, the expression of E-cadherin, α- and β-catenin and CD44V 6 in normal epidermis and on BCC cells were investigated. A significantly reduced expression of α-catenin and CD44V 6 and a slightly reduced expression of E-cadherin on BCC cells were observed compared with the overlying epidermis. Immunoelectron microscopy was used to investigate whether the decreased expression of E-cadherin and CD44V 6 was due to either an absence or downregulation of specific membrane structures or due to an overall downregulation of these adhesion molecules in all membrane structures in BCC. E-cadherin and CD44V 6 were expressed in adherens junctions, desmosomes, and complex interdigitating membrane structures both in normal epidermis and in BCC. A quantitative analysis showed that only a percentage of desmosomes was stained. In addition, the effect of pro-inflammatory cytokines, such as interferon-γ (IFN-γ) and tumor necrosis factor-α (TNF-α), was investigated in biopsy specimens of normal skin and BCC, using a biopsy culture system and immunohistochemistry. The expression of E-cadherin and CD44V 6 was not significantly decreased after culturing BCC or normal skin biopsy specimens for 48 hours with or without recombinant human (rHu)IFN-γ or rHuTNF-α. It may be concluded that the decreased expression of both E-cadherin and CD44V 6, observed in light microscopy, was not attributable to the absence of specific specialized structures in BCC and most likely also not caused by downregulation by local cytokines, but rather by generic downregulation of both of these adhesion molecules during malignant transformation.

Leishmaniasis and Kaposi's Sarcoma in an HIV-Infected Patient

To the Editor: The presence of Kaposi sarcoma and leishmaniasis in the same cutaneous lesion, which has been observed in six cases (1-6), often reveals infection of peritumoral skin (1,3,4) (i.e., within the noninvolved areas immediately surrounding the abnormal portion of the skin biopsy). Subsequent to our previous report (4), we have seen two additional instances of leishmaniasis in perineoplastic dermis of Kaposi's sarcoma. A similar situation was described in the letter of Abajo et al. In contrast, the presence of Leishmania in clinically uninvolved skin (i.e., within a biopsy specimen taken distant from lesional skin) of human immunodeficiency virus (HIV)-infected patient with visceral leishmaniasis (VL) has not been looked for. As we mentioned in our article (4), only one patient had Leishmania in such "normal" skin documented with the technique of split-skin smear. However, with this procedure, a sharp distinction cannot be made between simple peripheral blood infestation of the dermis versus actual cutaneous involvement. Since our previous report (4), we have seen one additional case of "silent" leishmaniasis in two normal skin biopsy specimens of a HIV-infected patient with a prolonged fever, who had refused bone marrow aspiration. All cases demonstrating Leishmania in "normal" skin showed an extravascular pattern of infestation. This finding demonstrates, in HIV-infected patients with VL, a widespread involvement of skin by Leishmania, which differs from a simple peripheral blood dissemination. In comparison to bone marrow aspirate, the diagnostic value of peripheral blood smears for discovery of VL in HIV-infected subjects remain low (7). As with both marrow aspiration, normal results on skin biopsy could be useful in confirming the diagnosis of VL. On the other hand, in HIV-infected patients with Mediterranean VL the skin could act as a reservoir of infection and allow human-to-human transmission. Abajo et al. mention a new example of presumed "silent" leishmaniasis (E. F. Dandén, P. Penas, L. Rios, J. Fraga, J. Alvar, A. Garcia-Diez, unpublished observations). Although their report is not yet published, on the basis of their letter, two questions still remain to be answered. First, are their cases best categorized as cutaneous leishmaniasis or VL with a cutaneous involvement? The other unknown is whether or not the biopsy specimens demonstrating the "silent" leishmaniasis were obtained adjacent to or distant from the clinically apparent cutaneous lesion. C. Perrin, M.D. P. Del Giudice, M.D. B. Taillan, M.D. Y. Lefichoux, M.D. JF. Michiels, M.D. Departments of Pathology, Infectious Disease, Internal Medicine, and Parasitology Laboratory; University of Nice; Nice, France

Transcription of human herpesvirus-like agent (HHV-8) in Kaposi's sarcoma.

Recently, DNA sequences of what appear to be a unique human herpesvirus-like agent (HHV-8) have been detected in different types of Kaposi's sarcoma (KS) tumors (Chang, Y., E.C. Cesarman, M.S. Pessin, F. Lee, J.C. Culpepper, D.M. Knowles, and P.S. Moore. 1994. Science (Wash. DC). 266:1865-1869). To further elucidate the possibility that HHV-8 plays a role in the pathogenesis of KS, the expression of HHV-8 RNA was examined in fresh KS tissue specimens which were found to harbor HHV-8 DNA by polymerase chain reaction (PCR). The transcription of HHV-8 RNA was detected by RT-PCR in 26 of 29 specimens (89.7%) of the KS tumors including 2 of 3 CKS and 24 of 26 AIDS-KS. No positive signal was detected in eight biopsy specimens of normal skin from healthy donors. By Northern blot analysis, the expression of HHV-8 was detected in 2 of 10 KS tumors examined. Furthermore, the RNA transcripts were observed in endothelial cells lining the irregular vascular spaces and perivascular spindle-shaped cells histologically characteristic of KS in 2 out of 8 different KS specimens examined by in situ hybridization using an antisense probe specific of HHV-8. The detection of RNA expression of HHV-8 in KS tumors further supports the possible etiopathogenic role of this virus in the development of KS.

Kaposi's sarcoma-associated herpesvirus DNA sequences in prostate tissue and human semen.

Sequences of novel herpesvirus, Kaposi's sarcoma-associated herpesvirus (KSHV), have been indentified in Kaposi's sarcoma tissue, but it is not known whether the virus is transmitted by sexual contact. Using the polymerase chain reaction (PCR), we searched for KSHV DNA sequences in ejaculates from 43 healthy men and tissue from the urogenital tract or prostate of 100 immunocompetent adults. In an unblinded analysis, we identified KSHV DNA sequences in 2 of 20 tissue specimens from the urinary tract (10 percent; 15 men and 5 women), 3 of 46 specimens from the female genital tract (6.5 percent), 4 of 18 specimens from the glans or foreskin (22 percent), 7 of 16 specimens from the prostate (44 percent), and 30 or 33 ejaculates (91 percent). By contrast, such sequences were present in 1 of 18 samples of normal skin (5.5 percent) and 1 of 14 samples of peripheral-blood mononuclear cells (PBMCs; 7.1 percent). Ejaculates and PBMC samples from each of 10 study subjects were analyzed in a blinded, coded fashion, along with PBMCs and biopsy specimens of normal skin from 4 and 8 other patients, respectively. This analysis confirmed the presence of KSHV DNA sequences in semen. Viral DNA was not found in the sperm heads but was present in the fraction of the ejaculates that contained urothelial and other types of cells. Point mutations were found in PCR products amplified from both prostate tissue and sperm samples. KSHV infects a large proportion of healthy adults and is probably transmitted by sexual contact.

Aberrant bcl-2 protein expression provides a possible mechanism of neoplastic cell growth in cutaneous basal-cell carcinoma.

The bcl-2 proto-oncogene encodes a protein that protects cells from programmed cell death (apoptosis). High levels of this protein confer a growth advantage to neoplastic cells even in the absence of a high mitotic rate. This gene is involved in the interchromosomal 14;18 translocation, an abnormality present in more than two-thirds of follicular lymphomas and in about 25% of other non-Hodgkin's lymphomas of the lymph nodes. A recent study also demonstrated the presence of high levels of bcl-2 protein in solid tumors such as squamous cell carcinomas of the lung and related it to a better prognosis. We analyzed bcl-2 protein expression in 20 cases each of basal- and squamous-cell carcinoma and in 5 biopsy specimens of normal skin, using a monoclonal anti-bcl-2 protein antibody with a standard 3-step immunoperoxidase technique on routinely fixed, paraffin-embedded tissue sections. Normal skin showed positive staining of the majority of keratinocytes in the epidermal basal layer. Bcl-2 positivity was also observed within the outer root sheath and the mesenchymal cells of the follicular papillae, the clear cells of the eccrine glands, and in some melanocytes at the dermo-epidermal junction. Neoplastic cells in all cases of basal-cell carcinoma showed a positive cytoplasmic reaction for bcl-2. All biopsy specimens of squamous-cell carcinoma were negative. Expression of bcl-2 protein could also be observed in the majority of peri- and intratumoral lymphocytes in both basal-cell carcinoma and squamous-cell carcinoma.(ABSTRACT TRUNCATED AT 250 WORDS)

Localization of111Indium-labeled tumor infiltrating lymphocytes to tumor in patients receiving adoptive immunotherapy. Augmentation with cyclophosphamide and correlation with response

The adoptive transfer of interleukin-2 (IL-2)-cultured tumor infiltrating lymphocytes (TIL) can cause tumor regression in patients with metastatic melanoma. Thirty-eight patients with metastatic melanoma receiving high dose IL-2 and TIL were studied for the ability of autologous 111In-labeled TIL to localize to metastatic tumor deposits by gamma camera imaging and biopsy. Single bolus cyclophosphamide was administered 24-36 hours before TIL infusion in 27 treatment courses. Tumor localization by 111In-labeled TIL was seen by gamma camera imaging in 26 (68.4%) treatment courses. In a univariate analysis of factors influencing TIL traffic, cyclophosphamide administration was significantly associated with the ability to localize tumor by radionuclide imaging (P2 = 0.026). Twenty-one of 26 (80.8%) treatment courses given with cyclophosphamide demonstrated tumor localization, compared with only 5 of 12 (41.7%) treatment courses without cyclophosphamide. In addition, patients whose 111In-labeled TIL imaged their tumor received significantly more TIL than did those that did not (P2 = 0.0052). Biopsies revealed a greater accumulation of 111In in cutaneous tumors than in normal skin biopsy specimens (0.0021 and 0.0004% injectate/gram of tissue, respectively; P2 = < 0.001). The median tumor-to-normal-skin ratio of simultaneous biopsies was 5.0. Finally, 10 of 26 (38.5%) patients who had tumor localization by scan had a clinical response, whereas no responses were noted in 12 patients whose tumors were not imaged (P2 = 0.022). CONCLUSIONS. Localization in tumor may be important in the mechanism of TIL antitumor activity because no clinical responses were seen in patients who did not have their tumors imaged with 111In-TIL. Cyclophosphamide administration before TIL and IL-2 therapy and the administration of large numbers of TIL appear to improve the frequency of TIL localization to tumor.

Ultrastructural changes of the skin induced by human leukocyte elastase and cathepsin G.

Human neutrophil elastase and cathepsin G at a concentration of 10(-6) M were found to attack various substrates when normal skin biopsy specimens were incubated at 37 degrees C for 1 h with either of these enzymes. Elastase damaged primarily hemidesmosomes, leading to the epidermal cleavage from the dermis, whereas cathepsin G damaged the membrane structures. Both these neutral proteinases were highly specific to basal lamina of blood vessels. This indicates that neutrophil elastase and neutrophil cathepsin G may play different roles in various skin diseases related to enhanced activity and infiltration of neutrophils.

Detection of the Interferon-gamma-Induced Protein 10 in Psoriasiform Dermatitis of Acquired Immunodeficiency Syndrome

Interferon-gamma-induced protein 10 is a 10-kd protein produced by human keratinocytes following an exposure to interferon gamma. Keratinocytes within psoriatic plaques and within delayed-type hypersensitivity reactions have been shown to stain strongly with an affinity-purified rabbit antibody prepared against interferon-gamma-induced protein 10, suggesting a possible role for interferon gamma in the production of the lesions. A psoriasiform eruption has been seen in patients with acquired immunodeficiency syndrome (AIDS). Its severity appears to correlate with the degree of immunodeficiency in the early stages of AIDS. We stained 10 lesions of psoriasiform dermatitis of AIDS with the anti-interferon-gamma-induced protein 10 antibody using immunoperoxidase techniques. As controls, we studied 10 lesions of non-AIDS psoriasis, six lesions of seborrheic dermatitis with psoriasiform hyperplasia, one lesion of lichen simplex chronicus, and four biopsy specimens of normal skin from patients with AIDS. In addition, normal skin specimens taken from patients with AIDS and human immunodeficiency virus-negative patients at time of autopsy were examined. An identical, strong and diffuse staining pattern was seen in all cases of psoriasiform dermatitis of AIDS, non-AIDS psoriasis, seborrheic dermatitis, and lichen simplex chronicus. The specimens of normal skin showed only weak basal layer staining with anti-interferon-gamma-induced protein 10. Thus, the presence of interferon-gamma-induced protein 10 in keratinocytes was associated with psoriasiform hyperplasia and could be detected in both AIDS-associated and classic psoriasis.

A Study of Factor XHIa and MAC 387 Immunolabeling in Normal and Pathological Skin

Immunolabeling with two new antibodies, Factor XIIIa and MAC 387, has been studied in routinely processed biopsy specimens of normal skin, subcutaneous tissues, lymph nodes, and a variety of pathological conditions. These presumptive cell markers of the monocyte-macrophage lineage appear to label totally different and possibly mutually exclusive subsets of cells. In normal skin, Factor XIIIa labeled fixed dermal connective tissue cells, emphasizing their dendritic morphological appearance. Factor XIIIa expression in fibroproliferative conditions and spindle cell tumors varied: in particular, scars, keloids, and dermatofibrosarcomas do not label, whereas histiocytomas, fibrous papule of the nose, and atypical fibroxanthomas were well labeled. In inflammatory conditions, increased numbers of Factor XIIIa cells were found, but most macrophages and epithelioid and multinucleate cells did not label. In contrast, normal dermal connective tissue cells did not label with MAC 387, nor did the constituent cells of various fibroproliferative disorders. In inflammatory conditions, variable numbers of MAC 387-positive cells were observed, corresponding to histiocytes and macrophages, but labeling of epithelioid cells and multinucleate foreign body giant cells was variable. Histiocytosis X cells do not express either label. The theoretical and practical implications of these results are explored.

A simple method for elution of IgA deposits from the skin of patients with dermatitis herpetiformis.

To better understand the role of autoimmunity in the pathogenesis of dermatitis herpetiformis, linear IgA bullous dermatosis or other skin disorders, the antigenic specificity of the immune reactants bound in vivo in the skin must be identified. In order to do so, one must first be able to elute these immune reactants from the skin. We describe here a simple method of eluting not only specifically bound IgG, but also IgA and other immunoglobulins and complement components from skin biopsy material. The method involves cutaneous washing of the entrapped serum proteins in PBS pH 7 and pH 5 buffers followed by specific immunoglobulin elutions at pH 3 and 2. The IgA deposits which could not be removed by this treatment were eluted by a combination of low pH (0.5 M citrate pH 2) and a chaotropic agent (2 M NaCl). The relative concentration of IgA in eluates when quantitated by fluoroimmunoassay were three- to five-fold higher in dermatitis herpetiformis skin biopsy specimens, than in eluates of bullous pemphigoid or normal skin biopsy specimens.