Monoclonal antibodies to various morphologic components of human skin

Somatic cell hybrids were established from the mouse myeloma, P3x63Ag8.653 cells, and the spleen cells of a mouse hyperimmune to human epidermal cells. Indirect immunofluorescence test with hybridoma culture fluids displayed that 253 out of 263 hybridoma cultures secreted antibodies reactive with the frozen sections of human skin. The hybridomas secreting unique antibodies to skin components were cloned and designated as AHS-1 to -8. Monoclonal antibodies (MoAb) from AHS-1, AHS-2, and AHS-3 hybridomas did detect cytoplasmic antigens present in the epidermal layer, eccrine ducts and glands (except MoAb AHS-1), and hair follicles. Enzyme-linked immunosorbent assay showed that the antigen recognized by either MoAb AHS-1 or MoAb AHS-2, but not by MoAb AHS-3, shares the antigenic determinant with antigen(s) present in keratin polypeptides isolated from human callus. MoAb AHS-4 to -8 distinctly stained each morphologic component of the skin tissues: cell membranes of epidermal cells by MoAb AHS-4, cytoplasms in upper layer cells of epidermis by MoAb AHS-5, dermis by MoAb AHS-6, basement membrane zone by MoAb AHS-7, and eccrine ducts by MoAb AHS-8.     

Monoclonal antibodies cross-reactive with group A streptococci and normal and psoriatic human skin

Infection with group A streptococci has been implicated as a factor capable of exacerbating psoriasis. In order to explore the possibility of cross-reactivity between streptococcal antigens and human skin in this phenomenon, skin from psoriatic patients and control subjects was reacted with 3 monoclonal antibodies against group A streptococci and antibody binding was estimated by the indirect immunofluorescence technique. Monoclonal antibody 54.2.8 stained the nuclei and cytoplasm of cells within the epidermis and epidermal appendages, as well as cells scattered throughout the dermis. In contrast, monoclonal antibodies 49.8.2 and 36.2.2 labeled the cytoplasm of epidermal cells and epidermal appendages but did not react with nuclei. No difference in the staining patterns of control skin and uninvolved skin from patients with psoriasis was observed. However, skin from psoriatic lesions contained large amounts of cross-reactive skin component(s). Sera from patients with guttate psoriasis did not react differently with normal or psoriatic skin when compared with normal sera. Western immunoblots of skin extracts demonstrated that monoclonal antibody 54.2.8 reacted with a family of proteins in the molecular weight range of 60-70K. The results indicate that component(s) in human skin share cross-reactive epitopes with group A streptococci. Immunologic cross-reactions between group A streptococci and human skin may play an important role in the exacerbation of certain skin disorders following streptococcal infections.     

Immunofluorescent localization of type I and III collagens in normal human skin with polyclonal and monoclonal antibodies

Anti-type I and type III collagen polyclonal antibodies and anti-type III collagen monoclonal antibodies were produced using type I and type III collagen extracted from the human placenta. An indirect immunofluorescence technique using these antibodies showed the same distribution of type III as well as type I collagen in the entire dermis of the normal human skin in nearly identical patterns. Previous immunofluorescent study indicated that type III collagen is present predominantly in the papillary dermis. However, our observation that monoclonal antibody recognizing the helical portion of type III collagen reacted with the entire thickness of the dermis which suggested the presence of type I and III collagen in close proximity in the whole thickness of normal human dermis.     

Immune response to gamma-irradiated injectable human amnion and human skin collagens in the rat

The immune response in rats to gamma-irradiated human amnion and human skin collagen was characterized through histologic and immunologic methods. Pepsin-extracted human amnion collagen and skin collagen were purified and reconstituted. Implants of amnion collagen demonstrated greater persistence than skin collagen. For amnion collagen implants, no significant inflammatory response was found. Fibroblast and adipocyte ingrowth and neovascularization were present. Conversely, obvious inflammatory infiltration was evident in the skin collagen implants. Enzyme-linked immunosorbent assay results showed that anti-amnion collagen antibody levels were significantly lower than anti-skin collagen antibody levels against their respective implant materials. The ratios of type I to type III collagen are 56:44 and 95:5 for amnion collagen and skin collagen, respectively. These findings suggest that in this heterologous type system, type III collagen-rich amnion collagen preparations appear superior to skin collagen for soft-tissue augmentation.     

Monoclonal antibodies in the study of malignant skin lymphomas

Comprehensive analysis of skin infiltrate cells, carried out in malignant lymphomas of the skin, has included examinations with the use of monoclonal antibodies (MCA). Basing on the clinical, histologic, and immunologic findings, mycosis fungoides has been diagnosed in 9 patients and T-cellular lymphoblastic lymphosarcoma consisting of T-suppressors in 1. The findings evidence that (1) mycosis fungoides is a tumor consisting of OKT4 cells in the majority of cases; (2) mature T-cell markers are sometimes lost in the course of mycosis fungoides, this being parallelled by the emergence of antigens characteristic of earlier differentiation stages, e. g. corticothymocytic ones; (3) immunologic heterogeneity is a characteristic feature of malignant lymphomas of the skin; this may be explained by both: the tumor progress of the clone and the therapeutic pathomorphosis; (4) use of MCA to a wide spectrum of leukocytic differentiation antigens permits an accurate estimation of the nature of the tumor cells and of the cellular relationships; this helps understand the pathogenesis of malignant lymphomas of the skin and develop new treatment modalities.     

Reproducibility of the immunofluorescent test for antimelanoma antibodies.

The reproducibility of the indirect immunofluorescent test for antibodies to human malignant melanoma cytoplasmic antigens was investigated by testing panels of melanoma and normal sera against cells of each of several different melanomas. Tests were performed in replicate, read blindly by two observers, and repeated on different days. Different observers agreed in the interpretation of replicate assays in approximately 80 to 90% of cases. Variability increased considerably when assays were repeated on different days or when different in as few as 21% of tests. Thus, the results of the indirect immunofluorescent test for melanoma cytoplasmic antibodies must, at the present time, be interpreted with great caution.     

Monoclonal antibodies for epidermal population analysis

Three keratin antibodies (RKSE 60, Clone 77, and a rabbit polyclonal) and 2 vimentin antibodies (Vim ab and a rabbit polyclonal) were investigated using frozen sections of normal and psoriatic skin. Of these, the monoclonals RKSE 60 and Vim ab were selected for quantitative population analysis of healthy epidermis, psoriatic uninvolved epidermis, and psoriatic lesions. Suspensions of isolated cells were prepared from biopsy specimens by trypsinization, and stained with RKSE 60 or Vim ab using an indirect immunofluorescence assay. Our results showed an increase in the germinative fraction from the normal value of 30% to almost 50% in the psoriatic lesion; in absolute terms this corresponds to a 6-fold increase in the size of the germinative compartment. More interesting, the germinative psoriatic uninvolved epidermis (38%) was also significantly higher than normal. The percentage of vimentin-positive cells (Langerhans cells and melanocytes) was nearly double that of normal in both the lesion and the uninvolved psoriatic epidermis. We conclude that, in contrast to statements frequently encountered in the literature, the "uninvolved" skin of the patient is morphologically and functionally different from that of the healthy individual.     

Analysis of the age-related composition of human skin collagen and collagens synthesized by fibroblast culture

Age-related differences in the composition and the post-translational modifications of human skin collagens were examined in the present study. The data were compared with results of collagen synthesis from in vivo-aged fibroblasts in culture. Skin extracts and newly synthesized collagen from fibroblast cultures derived from both old and young donor groups showed the same ratio of collagen III to collagen I. Furthermore, no difference was noted in the degree of prolyl and lysyl hydroxylation of collagen I and collagen III. Young and old fibroblasts synthesized a similar quantity of collagen in vitro. The data suggest that fibroblasts maintain a uniform level of collagen production, composition and modification independent of the age of the donor.     

Diagnostic value of antinuclear antibodies with special reference to the immunofluorescent speckled pattern

This article reviews the different immunofluorescence pattern of antinuclear antibodies (ANA) in various autoimmune diseases. Particular attention has been payed to the speckled immunofluorescence pattern, which is often caused by extractable nuclear antibodies (ENA). Clinical relevance, diagnostic significance and prognostic value are discussed in detail.     

Monoclonal anti-interleukin 2 (15-2) antibody binding to granular layer keratinocytes of human skin

Among several monoclonal antibodies (moABs) directed against human interleukin 2 (IL-2), the 15-2 moAB raised in our laboratory against unglycosylated recombinant IL-2 (produced in Escherichia coli) cross-reacted with a human skin epitope. This moAB gave a strong staining on the cell-surface membranes of keratinocytes from the granular layer of the epidermis. In addition, the 15-2 moAB stained 15% of epidermal cell suspensions obtained from suction blisters and reacted with cells from the spinous layer in parakeratosis and psoriasis, as well as with spinous epithelioma cells. Preincubation of the 15-2 moAB with pure human recombinant IL-2 abrogated skin binding, whereas a polyclonal antikeratin antiserum did not block 15-2 skin binding. Two other anti-IL-2 moABs, one directed against unglycosylated recombinant IL-2 (17-2 moAB) and one against glycosylated natural IL-2 (9B11 IE5 moAB), were unreactive on skin. Taken together, the data suggest that the 15-2 moAB binds to an epitope cross-reacting with, but different from, IL-2 which is located in the cell-surface membranes of granular layer cells. This cross-reactive epitope may provide a useful probe for the study of human epidermal cell differentiation.     

Wheat grain immunofluorescent antibodies as an indication of gluten sensitivity?

An immunofluorescence method using whole sections of wheat grains as the substrate was applied to detect circulating antibodies to wheat gluten in dermatitis herpetiformis patients and in controls. Only IgG class antibodies were detected. From dermatitis herpetiformis patients 22% had these antibodies as had 22% of the atopic dermatitis group. Among the controls who had no skin problems 12% were faintly positive. It is evident that the test as such is non-specific and does not have diagnostic significance in dermatitis herpetiformis.     

Demonstration of ß-lipoproteins in the psoriatic skin by immunofluorescent technique

Summary The immunofluorescent staining of lipoproteins in psoriatic skin was carried out using rabbit antiserum to human-lipoproteins. In active psoriatic lesions from 17 patients including 4 patients with psoriasis guttata acuta and a patient with psoriasis pustulosa (Zumbusch), the specific fluorescence suggesting increased deposits of-lipoproteins was constantly observed in the intercellular spaces of the epidermis and around or on the blood vessels in all layers of the corium. With the exception of pustulosis palmaris et plantaris (Lever), in which similar staining results were seen, the above-mentioned finding seems to be considerably characteristic of psoriasis. On the other hand, in 7 improved lesions and in 7 uninvolved regions, the epidermal fluorescence was remarkably diminished and tended to be localized or observed not at all, while the vascular fluorescence of the corium was hardly changed in 5 of 7 cases respectively.

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Zusammenfassung Die Lipoproteine psoriatischer Hauttypen wurden durch Immunfluorescenz unter Verwendung von Kaninchen-Antihumanbetalipoprotein-Antiserum dargestellt. In den aktiven Psoriasisherden von 17 Patienten sowie bei 4 Patienten mit Psoriais guttata akuta und einem weiteren Patienten mit Psoriasis pustulosa (Zumbusch) zeigte sich in den Intercellularräumen der Epidermis sowie über/um die Blutgefäße aller Dermisetagen eine spezifische Fluorscenz, die das Vorhandensein erhöhter Mengen von Betalipoprotein vermuten läßt. Mit Ausnahme der Pustulosis palmaris et plantaris (Lever), bei der ein ähnliches Reaktionsmuster beobachtet wurde, scheinen die hier mitgeteilten Resultate von kennzeichnender Bedeutung für die Psoriasis zu sein.Bei 7 abgeheilten Erkrankungsherden sowie in 7 nichterkrankten Hautarealen zeigte sich eine erheblich verminderte, teils lokal begrenzte oder auch fehlende epidermale Fluorescenz. Die an den Gefäßen lokalisierte Fluorescenz im Corium war jedoch bei 5 von 7 Patienten in beiden Fällen kaum vermindert.

     

Monoclonal antibodies selected to discriminate between malignant melanomas and nevocellular nevi

Two monoclonal antibodies (MoAbs), PAL-M1 and PAL-M2, are described that were selected to discriminate between melanomas and nevocellular nevi (NN) in frozen sections. MoAb PAL-M1 reacted with all 15 melanoma metastases (MM), with 14 of 19 primary cutaneous melanomas (PCM), 9 of 35 dysplastic nevi (DN), and 2 of 26 NN. The 2 NN stained were removed from patients with the dysplastic nevus syndrome. MoAb PAL-M2 reacted with 9 of 15 MM, 5 of 19 PCM, 3 of 35 DN, and did not react with 26 NN after usual staining conditions. The proportion of melanocytic cells stained was low in DN and much higher in PCM and especially in MM. Staining in DN was restricted to intraepidermal or subepidermal nests of atypical melanocytes. In PCM, staining with PAL-M2 was observed only in tumors with a Breslow thickness of 0.76 mm or higher. PAL-M1 and PAL-M2 may be immunohistochemical markers for tumor progression in melanocytic proliferations.     

122例皮肤直接免疫荧光试验的诊断价值探讨

本文报导应用荧光标记羊抗人lgG,IgA, IgM,C3、纤维蛋白原等对LE的皮损及正常皮肤、天疱疮、类天疱疮等皮损进行直接免疫荧光技术检查.结果证明直接免疫荧光技术对上述疾病具有一定的诊断价值.     

Antinuclear antibodies: a simplified classification of the nuclear immunofluorescent patterns.

The key to a simplified classification of the nuclear immunofluorescent patterns is to separate out only two patterns, the speckled and nucleolar, from the nonhomogeneous particulate group (showing stained particles). There are only six categories divided into two major groups: nonparticulate and particulate. The nonparticulate group consists of the (1) peripheral, (2) homogeneous, and (3) leukocyte specific patterns. The particulate group is divided into (1) nucleolar, (2) speckled, and (3) "other particulates." The major diagnostic and prognostic values of of the test are retained by the simple expedient of separating out only two morphologically distinct and diagnostically important patterns from the particulate group, the nucleolar and speckled patterns, seen mainly in scleroderma but not in lupus erythematosus.     

单克隆抗体治疗相关皮肤病的进展

利用单克隆抗体靶向病变组织或细胞上表面抗原已成为理想的靶向治疗方法。这种新方法受到大家的广泛关注,在皮肤病领域中也有着广阔的前景。