production and characterization of monoclonal antibodies to treponema pallidum

1
387 BLOOD COAGUUTION EXANINATION IN PROGRESIVE SYSTEHIC SCLEROSIS (PSS)---PLASMA LEVELS OF THROMBIN . ANTITHROMBIN LB COMPLEX(TAT) AND ANTITHROMBIN m (ATDI)--- MANABU MEDA, YUKIKO SHIKANO, AND SHUNJI NOR1 Department of Dermatology, Gifu University School of Medicine, We examined blood coagulation in 45 cases of PSS.especially fibrinogen@bg). prothrombin time(PT). active partial thromboplastin time(APTT), TAT, ATIII. plasninqen@ng). PDP, D-diner, crZ plasmin Inhibitor-plasmin complex(PICj. soluble fibrin monomer complex (SPMC) in order to clarify the disorder of platelet-blood vessel system. As the result. plasma levels of TAT and ATUl were high in 29 of 39 cases(74.4%), 23 of 45 cases(51.1%), respectively. There was no statistical negative correlation between those of TAT and ATlll. Plasma levels of PIC. D-dimer. FDP and SFNC were not always high except for several cases. and lnoreover there was no statistical positive correlation between those of TAT and PIC. These data lead us to recomfirm some disorder of platelet- blood vessel system in PSS. SERUM ADENOSINE DEAMINASE ACTIVITY IN SCLERODERNA PATIENTS TETSLJO SASAKI AND HIROSHI NAKAJIMA. Department of Dermatology, Yokohama City University School of Medicine, Yokohama, Japan Adenosine deaminase (ADA) is one of the enzymes of the purine metabolic pathway. In serum, ADA activity is mainly derived from T lymphocytes.The purpose of this study is to clarify the significanceof ADA in scleroderma.Serum ADA activity was determined with enzymatic method in 28 patients with systemic scleroderma (PSS), 4 localized scleroderma,4 MCTD, 6 dermaromyosiris (DM) and 14 with other related diseases, 53 (95 X) of which had antinuclear antibodies. Serum ADA activity was elevated in 89 4,of PSS, all of localized scleroderma and MCTD, 83 % of DM and 71 % of the other related diseases. The mean values in 9 PSS with anti Scl-70 antibodies, 6 PSS with anti RNP antibodies and 4 MCTD, and 8 PSS and I generalizedmorphea with anticentromere antibodies were 26.7, 25.6 and 22.2 IlJ/l, respectively.These results suggest that serum ADA activity may well reflect T cell activation in scleroderma and related disorders. SERUM SCSB-9 LEVELS AS A SENSITIVE INDICATOR OF HENOCW-SCHONLEINPURPURA ACTIVITY SEIJI KAWANA, YUSUKE SUZUKI'/, AND TOSHIYA ASAIII. Department of Dermatology, ST. Luke's InternationalHospital, Tokyo, Kitasato Univ. School of Medicine, Sagamiharail The concentrationof the terminal complement complex (TCC), SC5b-9, was determined for 48 serum samples from 30 patients with Henoch-Schonleinpurpura. In 25 of the patients, TCC concentrationwas found to increase significantly in the ac- tive phase of the disease. Three patients with various mani- festations of the disease were followed longitudinallyfor a period of several years covering in active and inactive pha- ses. TCC elevation in all cases was noted to be correlated with exacerbationsof the disease. C3 and C4 or CH50 levels in many instances remained normal or increased and thus were not reliable indicators of disease activity. Measurement of TCC should thus prove quite useful for monitoring the acti- vity of Henoch-Schonleinpurpura in patients in whom comple- ment activation is pathognomonic. PRODUCTION AND CHARACTERIZATION OF MONOCLONAL ANTIBODIES TO TREPONEMA PALLIDUM Min Geol Lee, Kyu Kwang Whang, Wook Lew. Jung Bock Lee Department of Dermatology. Yonsei University College of Medicine, Seoul, Korea Murine monoclonal antibodies against Treponema pallidurn were iwlated by measuring optical denshies or antibody tfters by enzyme-linked immunosorbent assay, indirect immunofluorescence or hemagglutination techmques. The isotypes of the 7 monoclinal antibodies produced were ; IgGl. lgG2a and IgGPb. Six of the seven monoclonal antibodtes were directed specifically against T. paltidum antigen. Five of the six monoclinal antibodies were raised against a polypeptide of a molecular weight of 47 kDa. One monoclonal antibody was bound to T. pallidurn, T phagedenis, T. denticola and T. vincentb, thus demonstrating a possible specificity for a treponema group antigen. This was raised against a polypeptide of a molecular weight of 64 kDa ELISA-inhibition tests wlth these monoclonal antibodies showed that there was a statistically significant difference in the degree of inhibition between groups of healthy controls and of each stage of syphilis. Therefore, the specific anttbodies reacting with major antigenic polypeptides of a molecular weight of 47 kDa could be adapted in the diagwxis 01 syphilis CO-EXPRESSION OF FILACCRIN AND TRICHOHYALIN IN NORMAL AND ABNORMAL KERATINIZED EPITHELIUM MOTOMU NANABE” ‘, W.MICHAEL O’GUIN”. BEVEILY A. DALE,‘, TUNGTIEN SUN’, JUNTENDO UNIVERSITY, TOKYO’, NEWYORK UNIV., NEW YORK”, AND UNIV. OF WASHINGTON, SEATTLE ‘. Keratohyalin and trlchohyalin granules are classically thouaht to retresent differentiation markers uniaue tu the epidermis and’ inner root sheath/ medulla of hair ‘foil icles, respectively. Using antinbodies specific for the protein subunits of these two types of granules. we discovered important exceptions to this rule. Keratohyalln and trichohyalin proteins co-exist in 1) papilla of the dorsal tongue epithelium and 2) bullous congenital ichthyosiform erythroderma and psoriasis. Furthermore, on the basis of ultrastructural double labellinn. we are now able to show that keratohyalin and trichohyaiin proteins are topological segregated in the same granuies forming hybrid granules. Thus provided evidence that keratohyalin and trichohyalin proteins undergo highly coordinated pathways of synthesis,, during the terminal differentiation of normal tongue eplthelium and some abnormal keratinlzed epithelium. A NEW MONOCLONAL ANTIBODY AGAINST HEMIDESMOSOMES OF HUMAN EPIDERMAL BASAL CELLS XIAO-MIN ZHANG, YUJI HORIGUCHI, SADAO IMAMURA. Department of Dermatology, Faculty of Medicine, Kyoto University, Kyoto Using monoclonal antibody technology,a new antibasement membrane zone (BMZ) antibody, designated as 2B7B has been derived from human prostate gland. Immunohistochemical ass& ravealed rhat this antigen is expressed not only along BMZ of the acini of prosfate gland but also along BMZ of the epidermis, appendages and epithelium of the esophagus, urinary bladder and ureter, as well as endorhelial cells of blood vessel. From the dermatologicalpoint, this antibody is interestingas it reacts similarly to the bullous pemphigoid antibodies to the hemidesmosomesof the epidermal base1 cell, though it does not compete the BP antibodies. This antibody seems much available to dermatoiogical investigation,such as the diagnosis of squamous cell carcinoma, basal cell epirhelioma and skin appendage tumors.

Upload: phungtuong

Post on 03-Jan-2017

218 views

Category:

Documents


2 download

TRANSCRIPT

387

BLOOD COAGUUTION EXANINATION IN PROGRESIVE SYSTEHIC SCLEROSIS

(PSS)---PLASMA LEVELS OF THROMBIN . ANTITHROMBIN LB COMPLEX(TAT)

AND ANTITHROMBIN m (ATDI)---

MANABU MEDA, YUKIKO SHIKANO, AND SHUNJI NOR1 Department of Dermatology, Gifu University School of Medicine,

We examined blood coagulation in 45 cases of PSS.especially fibrinogen@bg). prothrombin time(PT). active partial thromboplastin time(APTT), TAT, ATIII. plasninqen@ng). PDP, D-diner, crZ plasmin Inhibitor-plasmin complex(PICj. soluble fibrin monomer complex (SPMC) in order to clarify the disorder of platelet-blood vessel system. As the result. plasma levels of TAT and ATUl were high in 29 of 39 cases(74.4%), 23 of 45 cases(51.1%), respectively. There was no statistical negative correlation between those of TAT and ATlll. Plasma levels of PIC. D-dimer. FDP and SFNC were not always

high except for several cases. and lnoreover there was no statistical positive correlation between those of TAT and PIC.

These data lead us to recomfirm some disorder of platelet- blood vessel system in PSS.

SERUM ADENOSINE DEAMINASE ACTIVITY IN SCLERODERNA PATIENTS

TETSLJO SASAKI AND HIROSHI NAKAJIMA. Department of Dermatology, Yokohama City University School of Medicine, Yokohama, Japan

Adenosine deaminase (ADA) is one of the enzymes of the purine metabolic pathway. In serum, ADA activity is mainly derived from T lymphocytes. The purpose of this study is to clarify the significance of ADA in scleroderma. Serum ADA activity was determined with enzymatic method in 28 patients with systemic scleroderma (PSS), 4 localized scleroderma, 4 MCTD, 6 dermaromyosiris (DM) and 14 with other related diseases, 53 (95 X) of which had antinuclear antibodies. Serum ADA activity was elevated in 89 4, of PSS, all of localized scleroderma and MCTD, 83 % of DM and 71 % of the other related diseases. The mean values in 9 PSS with anti Scl-70 antibodies, 6 PSS with anti RNP antibodies and 4 MCTD, and 8 PSS and I generalized morphea with anticentromere antibodies were 26.7, 25.6 and 22.2 IlJ/l, respectively. These results suggest that serum ADA activity may well reflect T cell activation in scleroderma and related disorders.

SERUM SCSB-9 LEVELS AS A SENSITIVE INDICATOR OF HENOCW-SCHONLEIN PURPURA ACTIVITY

SEIJI KAWANA, YUSUKE SUZUKI'/, AND TOSHIYA ASAIII. Department of Dermatology, ST. Luke's International Hospital, Tokyo, Kitasato Univ. School of Medicine, Sagamiharail

The concentration of the terminal complement complex (TCC), SC5b-9, was determined for 48 serum samples from 30 patients with Henoch-Schonlein purpura. In 25 of the patients, TCC concentration was found to increase significantly in the ac- tive phase of the disease. Three patients with various mani- festations of the disease were followed longitudinally for a period of several years covering in active and inactive pha- ses. TCC elevation in all cases was noted to be correlated with exacerbations of the disease. C3 and C4 or CH50 levels in many instances remained normal or increased and thus were not reliable indicators of disease activity. Measurement of TCC should thus prove quite useful for monitoring the acti- vity of Henoch-Schonlein purpura in patients in whom comple- ment activation is pathognomonic.

PRODUCTION AND CHARACTERIZATION OF MONOCLONAL ANTIBODIES TO TREPONEMA PALLIDUM

Min Geol Lee, Kyu Kwang Whang, Wook Lew. Jung Bock Lee Department of Dermatology. Yonsei University College of Medicine, Seoul, Korea

Murine monoclonal antibodies against Treponema pallidurn were iwlated by measuring optical denshies or antibody tfters by enzyme-linked immunosorbent assay, indirect immunofluorescence or hemagglutination techmques. The isotypes of the 7 monoclinal antibodies produced were ; IgGl. lgG2a and IgGPb. Six of the seven monoclonal antibodtes were directed specifically against T. paltidum antigen. Five of the six monoclinal antibodies were raised against a polypeptide of a molecular weight of 47 kDa. One monoclonal antibody was bound to T. pallidurn, T phagedenis, T. denticola and T. vincentb, thus demonstrating a possible specificity for a treponema group antigen. This was raised against a polypeptide of a molecular weight of 64 kDa ELISA-inhibition tests wlth these monoclonal antibodies showed that there was a statistically significant difference in the degree of inhibition between groups of healthy controls and of each stage of syphilis. Therefore, the specific anttbodies reacting with major antigenic polypeptides of a molecular weight of 47 kDa could be adapted in the diagwxis 01 syphilis

CO-EXPRESSION OF FILACCRIN AND TRICHOHYALIN IN NORMAL AND ABNORMAL KERATINIZED EPITHELIUM

MOTOMU NANABE” ‘, W.MICHAEL O’GUIN”. BEVEILY A. DALE,‘, TUNGTIEN SUN’, JUNTENDO UNIVERSITY, TOKYO’, NEW YORK UNIV., NEW YORK”, AND UNIV. OF WASHINGTON, SEATTLE ‘.

Keratohyalin and trlchohyalin granules are classically thouaht to retresent differentiation markers uniaue tu the epidermis and’ inner root sheath/ medulla of hair ‘foil icles, respectively. Using antinbodies specific for the protein subunits of these two types of granules. we discovered important exceptions to this rule. Keratohyalln and trichohyalin proteins co-exist in 1) papilla of the dorsal tongue epithelium and 2) bullous congenital ichthyosiform erythroderma and psoriasis. Furthermore, on the basis of ultrastructural double labellinn. we are now able to show that keratohyalin and trichohyaiin proteins are topological segregated in the same granuies forming hybrid granules. Thus provided evidence that keratohyalin and trichohyalin proteins undergo highly coordinated pathways of synthesis,, during the terminal differentiation of normal tongue eplthelium and some abnormal keratinlzed epithelium.

A NEW MONOCLONAL ANTIBODY AGAINST HEMIDESMOSOMES OF HUMAN EPIDERMAL BASAL CELLS

XIAO-MIN ZHANG, YUJI HORIGUCHI, SADAO IMAMURA. Department of Dermatology, Faculty of Medicine, Kyoto University, Kyoto Using monoclonal antibody technology, a new antibasement membrane zone (BMZ) antibody, designated as 2B7B has been derived from human prostate gland. Immunohistochemical ass& ravealed rhat this antigen is expressed not only along BMZ of the acini of prosfate gland but also along BMZ of the epidermis, appendages and epithelium of the esophagus, urinary bladder and ureter, as well as endorhelial cells of blood vessel. From the dermatological point, this antibody is interesting as it reacts similarly to the bullous pemphigoid antibodies to the hemidesmosomes of the epidermal base1 cell, though it does not compete the BP antibodies. This antibody seems much available to dermatoiogical investigation, such as the diagnosis of squamous cell carcinoma, basal cell epirhelioma and skin appendage tumors.