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Biomedical subjects

J C Huff

Publications and source records attributed to J C Huff.

At least 55 records · Page 3Linked to original sources
PubMed1987

An immunopathological study of herpes-associated erythema multiforme.

Any pathogenetic mechanism proposed for erythema multiforme (EM) must account for the prominent mononuclear cell infiltrate in the skin lesions. The purpose of this study was to characterize immunopathologically, with monoclonal antibodies to human leukocyte antigens, the inflammatory cells in early target lesions of recurrent herpes-associated EM. Cryostat sections of snap-frozen skin biopsies were studied by the avidin-biotin immunoperoxidase technique with use of the following monoclonal antibodies: anti-HLA-DR, anti-Leu M5, anti-Leu 4 + 5b, anti-Leu 3a + 3b, anti-Leu 2a, anti-Leu 14, and anti-Leu 6. The dermal mononuclear inflammatory infiltrate in the EM biopsies consisted of monocyte-macrophages and T-lymphocytes, with both helper and suppressor T cells present. Both the dermal inflammatory infiltrate and the overlying keratinocytes were strongly HLA-DR positive. No definite alteration of Langerhans cell number or distribution was noted. These findings are consistent with the characteristics seen in cell-mediated immune reactions in the skin and point to this as a likely immune mechanism for the tissue damage of EM.

Adult↗
PubMed1986

Proliferating cells of human basal cell carcinoma are located on the periphery of tumor nodules.

Study of the growth characteristics of basal cell carcinoma (BCC), a relatively well-organized, slow-growing skin cancer, has been limited because of the lack of methods for propagation of the tumor off the human host. We have used newly developed techniques for transplantation and propagation of BCC on athymic mice to study [3H]thymidine incorporation by nodular BCC. In human BCCs labeled in vitro immediately after removal from the mice and in vivo on the mice, [3H]thymidine during a 4-h pulse was incorporated primarily by cells on the periphery of tumor nodules (labeling indices 6-24%) rather than by the cells more central in tumor nodules (labeling indices 0-2%). Similar results were also seen when samples of tumor freshly removed from patients were labeled in vitro. We conclude that the dividing cells within nodular BCC are primarily the cells at the edges of tumor nodules and that this characteristic is related to the slow, progressive, invasive growth of BCC.

Animals↗
PubMed1986

Study of HLA-DR synthesis in cultured human keratinocytes.

Within the normal human epidermis only Langerhans and indeterminate cells express HLA-DR. Human keratinocytes (HK), however, may also express HLA-DR in certain disease states characterized by mononuclear cell infiltrates. Previous studies have shown that HK synthesize HLA-DR in response to stimulation by interferon gamma (INF-gamma). The purposes of this study were to define conditions under which cultured HK might express HLA-DR and to compare the HLA-DR synthesis of HK with that of monocytes. HLA-DR expression by HK as determined by indirect immunofluorescence of HK cultures was absent under standard low calcium conditions and remained absent with the addition of calcium, serum, mitogens, and supernatants from Pam-212 cells containing epidermal thymocyte-activating factor. HLA-DR expression in HK was induced by cocultivation with concanavalin A-stimulated peripheral blood mononuclear cells (PBMC), but not unstimulated PBMC. This effect was time-dependent and directly related to the number of PBMC. HLA-DR expression was also induced in a time- and dose-dependent manner by addition of supernatant from stimulated PBMC (SS) or by addition of recombinant INF-gamma but not by addition of interleukin (IL)-1 or IL-2. Induction by either SS or INF-gamma was blocked by an antiserum to INF-gamma. As determined by a semiquantitative immunoprecipitation technique, HLA-DR synthesis by HK was directly related to INF-gamma concentration. The pattern of HLA-DR peptides produced by HK was similar to that of monocytes, but the relative quantity synthesized was far less than that of monocytes.

Cells, Cultured↗
PubMed1986

Basal cell carcinomas grown in nude mice produce and deposit fibronectin in the extracellular matrix.

Epidermal cells in vitro produce and deposit fibronectin (FN) in the pericellular matrix. Such FN production by epidermal cells may be involved in vivo in wound reepithelialization, tissue morphogenesis, and growth of epithelial tumors. The purpose of this study was to examine whether the FN, previously shown to be within and surrounding human basal cell carcinoma (BCC) lobules, was in part the product of epidermal-derived tumor cells. To examine this question we took advantage of our ability to grow human BCC in nude mice. Since we could demonstrate that all stromal cells surrounding the BCC were of mouse origin, antibodies specific for human FN would distinguish epithelial-derived FN from mesenchymal-derived FN. Five solid BCCs were implanted subcutaneously in nude mice. Growing tumors were removed after 60 days, snap-frozen, sectioned on a cryostat, and verified microscopically as BCC. The Hoescht DNA stain, which can distinguish mouse and human nuclei, demonstrated that mouse, not human, fibroblasts occupied the stroma surrounding each tumor lobule. Sections of all 5 BCCs were stained by immunofluorescence and immunoperoxidase techniques with antibodies to bullous pemphigoid (BP) antigen, laminin (LM), and FN. BP antigen and LM were present at the basement membrane zone (BMZ) of all tumor lobules as previously described for in situ BCC. FN staining was present along the BMZ, within the tumor lobules, and in the surrounding stroma. Antibodies to human FN were passed over a mouse FN affinity column to absorb antibodies which cross-reacted with mouse FN. The resultant antibody preparation, which was specific for human FN in this system, continued to demonstrate FN along the BMZ and within the tumors, but failed to stain FN in more distant stroma. Epidermal-derived cells, therefore, can synthesize and deposit FN in vivo in adjacent extracellular matrix. We speculate that this FN matrix may facilitate growth of BCC in this model.

Animals↗
PubMed1986

The biology of basal cell carcinomas--a revisit and recent developments.

This paper will review some of the present information on the biology of basal cell carcinoma. Recent developments include a better understanding of the basement membrane components surrounding the tumor nodules, as well as identifying the most actively dividing cells in solid basal cell carcinoma by autoradiographic techniques. The implications of a recently developed animal model will also be discussed.

Animals↗
PubMed1985

Transplantation of human basal cell carcinomas to athymic mice.

Basal cell carcinomas (BCCs) obtained from 22 subjects undergoing microscopically controlled surgery were transplanted to 40 athymic (nude) mice. With no further immunosuppression of the mice, no tumor growth was noted in the first 14 attempts. When mice were further immunosuppressed with anti-lymphocyte serum (ALS) injections and by splenectomy, successful tumor growth was achieved in 15 of 22 mice by a subcutaneous implantation technique and in 1 of 4 by a superficial grafting technique. Transplanted BCC retained the morphology and basement membrane proteins typical of human BCC. As determined by autoradiography, 3H-thymidine was incorporated primarily in the peripheral palisaded cells of the transplanted tumor. Successful use of the athymic mouse model for study of human BCC requires use of mice further immunosuppressed by splenectomy and ALS, and the use of a subcutaneous implantation technique. With the use of this model, studies of the biology of human BCC may be possible.

Animals↗
PubMed1985

Chronic graft-versus-host disease as a model for scleroderma. II. Mast cell depletion with deposition of immunoglobulins in the skin and fibrosis.

We explored the pathologic changes in the skin of mice undergoing a chronic graft-versus-host (GVH) reaction. In rodents and in man, chronic GVH includes the deposition of excess collagen in the skin-a reaction which resembles idiopathic scleroderma. GVH disease across minor histocompatibility barriers was produced by injecting B10.D2 cells into irradiated BALB/c mice. These strains are identical at the H-2 and Mls loci but differ in minor histocompatibility antigens. Control BALB/c mice received irradiation and BALB/c cells. Serial skin biopsies were taken and studied for histological changes characteristic of chronic GVHD, for mast cell density, and for the deposition of immunoreactants. GVHD was produced in B10.D2----BALB/c mice as measured by body weight loss and the production of skin changes including dermal fibrosis, loss of fat and appendages, and a mononuclear cell infiltrate. Dermal mast cells, assessed by toluidine blue staining, were normal at Day 11, but had disappeared by Days 21-63 and returned to normal by Day 104. Immunoglobulins IgG, IgA, and IgM appeared at the dermo-epidermal junction and along the basement membrane zone of hair follicles. This deposition was maximal at Day 42 and waned thereafter. Thus the appearance of immunoglobulins in the skin was maximal when mast cell staining was minimal. The changes in this GVHD model leading to a scleroderma-like picture in the skin are compatible with an immune etiology for the fibrosis. Vasodilation following liberation of mast cell mediators would facilitate the deposition of immunoglobulins. The disappearance of mast cell staining may be caused by extensive degranulation. We postulate an interaction between GVHD-activated T cells, mast cell stimulation, fibroblast activation, and fibrosis.

Animals↗
PubMed1985

Lymphedema of the lower abdominal wall in pregnancy.

Two patients with lymphedema of the lower abdominal wall in late pregnancy are presented. Prominent tortuous, dilated lymphatic channels were observed clinically. Mechanical obstruction of lymphatic drainage is thought to be the cause. Both patients improved after delivery.

Abdominal Muscles↗
PubMed1985

A simple and improved method for direct and indirect immunofluorescent staining.

Several technics for immunofluorescent staining have been described in the literature and are presently used by laboratories performing direct and indirect tissue immunofluorescence. The purpose of this communication is to describe a simple immunofluorescent technic that uses a five-slide polypropylene slide mailer containing the diluted conjugated antisera. This technic eliminates the need for an incubation chamber, decreases background staining, and uses less antiserum.

Fluorescent Antibody Technique↗
PubMed1985

The demonstration of SS-A/Ro antigen in human fetal tissues and in neonatal and adult skin.

Anti-SS-A/Ro antibodies have been statistically associated with congenital heart block and cutaneous lupus in the neonatal lupus syndrome, and with photosensitive cutaneous lupus in subacute cutaneous lupus erythematosus, but it had not been demonstrated that SS-A/Ro antigen is present in fetal tissues or at any age in human skin. We examined normal fetal tissues, normal neonatal and adult skin, and keratinocytes from purified serum-free cultures by immunofluorescence (IF) and by immunodiffusion or counterimmunoelectrophoresis (CIE) to determine the presence of SS-A/Ro antigen. We found SS-A/Ro antigen to be present in fetal hearts and in other internal organs. SS-A/Ro antigen could be demonstrated in biopsies of neonatal and adult skin by IF and was confirmed to be in keratinocytes by CIE. SS-A/Ro antigen may be found on the surface of keratinocytes in culture and therefore may be present at a relevant site for antibody binding. We have shown that SS-A/Ro antigen is a normal component of tissues that may be affected in neonatal lupus (fetal myocardium, neonatal epidermis) and in subacute cutaneous lupus erythematosus (adult epidermis).

Antibodies, Antinuclear↗
PubMed1985

Expression of fibronectin, laminin, and factor VIII-related antigen during development of the human cutaneous microvasculature.

The regulation of angiogenesis during human skin development is poorly understood. Since fibronectin is involved in cell movement and organization during embryogenesis and morphogenesis in a variety of species, we investigated the expression of fibronectin and factor VIII-related antigen, a marker for endothelial cells, at various stages in the development of the human cutaneous microvasculature. Skin specimens were obtained from 4 human fetuses during the second trimester (14-18 weeks), from newborn foreskins, and from consenting normal adults. Cryostat sections were stained with both fluorescein-conjugated rabbit antihuman fibronectin and rhodamine-conjugated goat antihuman factor VIII-related antigen. Expression of fibronectin in the microvasculature was striking in fetal skin but became progressively less prominent with maturation. Fibronectin appeared in fetal blood vessels as a bright continuous linear array, in neonatal blood vessels as a bright interrupted linear and speckled array, and in adult blood vessels as a sparse interrupted linear and speckled array. In contrast, expression of factor VIII-related antigen by the endothelium became more prominent with the degree of maturation of the microvasculature. Granular factor VIII-related antigen staining was scant in the newly forming blood vessels of fetal skin, bright but focal in the microvasculature of newborn skin, and intense and almost confluent in the blood vessels of adult skin. Although expression of fibronectin and factor VIII-related antigen changed, expression of laminin was consistent throughout development. Staining of the same skin specimens with fluorescein-conjugated sheep antihuman laminin produced a bright continuous linear pattern in all blood vessels. The reciprocal relationship manifested by intense fibronectin staining during human blood vessel development and prominent factor VIII-related antigen staining in mature blood vessels supports the hypotheses that fibronectin plays a role in human blood vessel modulation and morphogenesis, and that factor VIII-related antigen is a marker for endothelial cell differentiation.

Adult↗
PubMed1985

Subacute cutaneous lupus erythematosus associated with hydrochlorothiazide therapy.

Photosensitive eruptions with clinical and histologic features of subacute cutaneous lupus erythematosus and antibodies to SS-A(Ro) antigen occurred in five patients taking hydrochlorothiazide. After drug therapy was discontinued, the eruptions cleared. In one patient anti-SS-A antibodies disappeared after discontinuation of thiazide, and in another rechallenge with hydrochlorothiazide produced an acute dermatitis with a photodistribution. These eruptions may represent a new type of photosensitive drug reaction in which the photoactive drug may be synergistic with anti-SS-A antibody in producing cutaneous lesions of photosensitive subacute cutaneous lupus erythematosus.

Adult↗
PubMed1985

Erythema multiforme.

In summary, the diagnosis of erythema multiforme is appropriate for a self-limiting or episodic cutaneous or mucocutaneous illness with skin lesions morphologically and histologically compatible. With typical erythema multiforme minor, characterized by classic skin lesions with or without oral erosions, most patients' disease is associated with recurrent herpes simplex infections. This is particularly true with recurrent erythema multiforme. Symptomatic conservative care, antibiotic treatment for purulent secondarily infected oral lesions, and avoidance of systemic steroids are appropriate therapeutic guidelines. The more serious syndrome, erythema multiforme major, or Stevens-Johnson syndrome, is characterized by skin lesions that are somewhat atypical and different from those of erythema multiforme minor in association with erosions on multiple mucosal surfaces. Drugs and mycoplasmal infections are important precipitating factors for erythema multiforme major. Hospitalization and laboratory tests are often required because of the severity of the illness and the occasional damage to other organ systems. Conservative, symptomatic care, withdrawal of any drug that may have caused the illness, treatment of any mycoplasmal infection, and antibiotic therapy for purulent secondarily infected lesions are worthwhile therapeutic measures. Early treatment with systemic steroids may be helpful in preventing further damage, and the risks and potential benefits of such therapy must be evaluated on an individual basis.

Adrenal Cortex Hormones↗
PubMed1984

Erythema multiforme: clinical, histopathologic, and immunologic study.

In a prospective study of erythema multiforme, forty-two cases were selected with the use of defined criteria. In thirty-three cases (79%), the erythema multiforme occurred following a lesion of recurrent herpes simplex; in four cases (10%), it was related to administration of a sulfonamide drug. Herpes-associated erythema multiforme (HEM) was largely recurrent erythema multiforme minor and was characterized histopathologically by inflammatory changes, such as spongiosis and exocytosis, and by focal liquefaction degeneration of the basal cell zone of the epidermis. Sulfa-associated erythema multiforme (SEM) was a nonrecurrent illness with widespread cutaneous and mucosal damage associated with prominent histologic necrosis of epidermal cells. The deposition of C3 and fibrin along the dermoepidermal junction and the deposition of IgM, C3, and fibrin around dermal blood vessels by immunofluorescence microscopy were similar in both groups. Although HEM and SEM may have somewhat different clinical and histologic features, there is significant overlap in the pattern of tissue damage.

Adolescent↗
PubMed1984

Fibronectin in basal cell epithelioma: sources and significance.

Nodular basal cell epitheliomas (BCE) contain fibronectin both within tumor nodules and at the nodule-stroma interface (basement membrane zone). Fibronectin within or at the periphery of tumor nodules could be derived from the tumor cells, from entrapped stroma, or from plasma. The present study was designed to elucidate the source(s) of fibronectin within BCE nodules. If stromal entrapment occurred to any great extent, von Willebrand factor VIII:Ag-stained blood vessels within tumor nodules should be evident by immunofluorescence techniques. Likewise, if plasma proteins were deposited in BCE, the tumor nodules should stain with fluorescein-conjugated antifibrinogen antibodies. Therefore, 6 basal cell epitheliomas were double labeled with rhodamine-conjugated antihuman fibronectin and fluorescein-conjugated antihuman factor VIII:Ag or fluorescein-labeled antihuman fibrinogen. Fibronectin was present in a linear disposition along the margin of tumor lobules and as a fine filamentous deposit in the central portions of tumor tissue. There was no evidence of fibrinogen or factor VIII:Ag in any of the tumor lobules. Factor VIII:Ag was present in a granular pattern within blood vessel walls that coursed between tumor nests. An indirect immunoperoxidase technique using rabbit antihuman fibronectin and peroxidase-labeled goat antirabbit IgG demonstrated that fibronectin within the central portion of the tumor lobules was closely associated with the tumor cells. The absence of fibrinogen and factor VIII:Ag within the tumor tissue indicates that the fibronectin is probably not plasma- or stroma-derived while immunoperoxidase data suggest that fibronectin may be a product of BCE cells.

Antigens↗
PubMed1984

Ultraviolet light induces binding of antibodies to selected nuclear antigens on cultured human keratinocytes.

Antibodies which bind to different nuclear antigens in tissue sections or in permeabilized cell cultures are useful markers of subsets of connective tissue disease, especially of lupus erythematosus (LE), but whether these antibodies are able to react with these intracellular sequestered antigens in vivo and cause immunologic tissue damage has been a matter of much debate. We report experiments which show that ultraviolet light-irradiated, cultured human keratinocytes bind IgG antibodies from the sera of LE patients with either monospecific anti-SSA/Ro, anti-RNP, or anti-Sm activity, which implies that these antigens have been made accessible on the cell surface by ultraviolet irradiation. Normal human sera or LE patient's sera with anti-double-stranded DNA, anti-single-stranded DNA, or antihistone activity do not bind to the surface of irradiated human keratinocytes. In control experiments, all antisera produced the expected patterns of nuclear and cytoplasmic staining of fixed permeabilized, unirradiated keratinocytes. Careful study of the viability and permeability of irradiated keratinocytes by several techniques showed that this apparent cell membrane expression of extractable nuclear antigens (SSA/Ro, RNP, and Sm) following irradiation was seen on injured keratinocytes whose cell membranes were intact, but not on dead cells. It is particularly significant that all six monospecific anti-SSA/Ro sera bound to irradiated keratinocytes, since this antibody antigen system is highly associated with photosensitive cutaneous LE.

Antibodies, Antinuclear↗