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나환자의 혈청내 Immunoglobulins, a2-macroglobulin 및 보체 (C3)량에 관한 연구

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 Determination of serum immunoglobulins, a2-macroglobulin and complement (C3) levels in leprosy patients 
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[한글] 나병에 있어서의 면역기전과 나반응에 과해서는 많은 보고가있다. 나종양형의 경우 체 액성면역은 항진되어있으나 세포성면역은 정상인 또는 결핵양형에서 보다 저하되어 있는 것으로 알려져있다. 체액성면역의 항진을 뒷받침하는 실험으로 나종양형 환자에서 그 혈 청내 globulin량이 전체적으로 증가되어 있다는 점에 대해서는 여러 연구자 간에 별다른 이론이 없다. 그러나 각기의 immunoglobulin olass의 혈청내 농도에 관해서는 보고자에 따라 상당한 차이를 나타내고 있으며, 이러한 차이는 immunoglobulin량의 측정방법 및 검 사 대상자의 선정 여하에 따라 야기되는 것ㅇ로 보고되어있다. 만흥 나학자들은 나반응을 immune complex disease의 일종으로 간주하게 되었으며 나성결절성홍반(羅性結節性紅斑: Erythema Nodosum Leprosum)의 피부환부증상은 Arthus 현상으로 풀이하였으며, 또 나병 에서 특히 나반응의 경우 그 혈청내에서 자가면역양요소(自家免疫樣要素: Autoimmune lik e factors)의 검출 및 매독혈청검사에 대한 고율의 생물학적 위양성등 소위 자가면역기전 (autoimmune mechanise)에 의한 것임을 입증하려는 보고가 있다. 이에 저자는 나종야형 환자에 있어서의 체액성면역 항진의 본체 및 나반응발생의 기전 을 추구하기위한 연구의 일환으로 혈청내 immunoglobuline (IgG, IgM 및 IgA), α^^2 -ma croglobulin 및 보체(C3)량을 측정하고 동시에 VDRL(Venereal Disearch Laboratory) slid e 정량시험 및 C-reactive protein 시험을 실시하여 다음과 같은 결과를 얻었다. 1. 나종양형 환자 혈청내 IgG 및 IgA 량은 정상 대조군에 비하여 통계학적으로 의의있 는 증가(p<0.001, p<0.01)를 보였으나 IgM의 양은 증가되어 있었으나 통계학적 유의치(p> 0.05)는 되지 못하였다. 2. α^^2 -macroglobulin 량은 나종양형 환자군, 결핵양형 환자군 및 정상 대조군간에 서 차이를 볼 수 없었으나 나성결절성홍반(ENK) 병력이 있었던 환자군은 ENL 병력이 없었 던 환자군에 비하여 통계학적으로 의의있게 )p<0.05) 저하되어 있었다. 3. 보체 (C3)량은 나종양형 환자군에서 정상 대조군 및 결핵양형 환자군에 비하여 현저 히 증가되어 있는것(p<0.02, p<0.00)을 볼 수 있었으며, 피부조직(Wade's skin scraping method)에서의 세균지수가 2+이하인 환자군보다는 2+이상인 환자군에서, 또한 ENL 병력이 없었던 환자군(124±27.6mg/100ml) 보다는 ENL병력이 없었던 환자군 (141±21.7mg/100ml )에서 고가의 혈청내 보체(C3)량이 측정되었다. 4. C - reactive protein 시험결과 나종양형 환자 38례중 10례 (26.3%)에서, ENL병력이 있었던 나종양형 환자 9례중 5례(55.6%), ENL 병력이 없었던 나종양형환자 29례중 5례(1 7.8%)에서 각각 양성을 나타났었으나 결핵양형 환자 10례에서는 모두 음성이었다. 5. VDRL(Venereal Disease Research Laboratory) slide 정량 시험에서 나종양형 환자 2 8례중 3례 (7.9%)에서 각각 1:32 1:4, 및 1:2의 응집가를 나타내었다.
[영문] Immune mechanism of leprosy is one of the obscure problems in the field of leprology. Studies of immune mechanism in leprosy have indicated that lepromatous leprosy patients suffer from hypoactive state of cell mediated immunity (Bullock, 1968; Turk, 1969; Turk and Waters, 1969; Godal et al., 1971; Hans et al., 1971; Mendes et al., 1974) and from hyperactive state of humoral immunity (Almeida, 1970), but in tuberculoid leprosy cell mediated immunity is normal and the humoral immunity is normal or slightly increased. Meulling et al. (1960) and Mathews Trautman (1965) observed the abnormal increase of serum globulin level in the sera from the patients with lepromatous leprosy by chemical method and paper electrophoresis and Lim and Fusaro (1964) detected the increase of gamma globulin by the method of immunoelectrophoresis. Following the report of Sheagren et al. (1969) that serum IgG and IgA levels in the patients with lepromatous leprosy were higher than that of normal control group, a number of study has been made on the serum concentration of each immunoglobulin class in the sera from the patients with lepromatius leprosy. Bullock et al. (1970) found that IgG, IgA and IgM levels in the lepromatious leprosy and normal control groups, but Fusaro (1970) observed the higher levels of serum IgG and IgM, and Malaviya et al. (1972) reported that only serum IgA in lepromatous leprosy sera was increased significantly. Kunal and Mittal (1972) showed that the IgA and IgM levels in the sera from lepromatous leprosy patients were higher comparing with those of tuberculoid leprosy patients were higher comparing with those of tuberculoid leprosy patients and normal control group, but IgG level was normal. ENL (erythema nodosum leprosum) is particularly common in the course of the treatment of lepromatous leprosy patients, in which more than 50 percent of these patients suffer from ENL by the end of the first year of treatment. It has been widely assumed that the mechanism of ENL have immunological bases. Turk and Waters (1969), Wemambu et al. (1969) and Waters et al. (1971) demonstrated the deposit of immune complex in the skin lesion of ENL by immunofluorescent studies, and suggested that ENL is a clinical manifestation of Arthus phenomenon. Azevedo and Melo (1966) reported that complement level in the sera from the patients with lepra reaction was lower than that of normal control group. However, Sheagren et al. (1969), Wemambu et al. (1969), and Mayama (1971) showed that serum complement level is higher in the patients with lepra reaction. The hypothesis that an autoimmune mechanism has a sole on the immunological responses in leprosy have attracted attentions of many research workers. A wide spectrum of autoimmunelike factors has been found in leprosy, including rheumatoid factor (Cathcart et al., 1961), thyroglobulin antibody (Bonomo et al., 1963), cold-precipitated proteins (Mathews and Trautman, 1965), as well as biological false positive in serological tests for syphilis (Edmundson et al. 1954; Ruge, 1955; Kim and Lew, 1965). Reports have been made on the presence of C-reactive protein in various inflammatory diseases, but the first one of the C-reactive protein in leprosy was made by Rabson (1955) who observed the higher frequency of C-reactive protein in the active lepromatous leprosy cases. His finding was amply confirmed by others who found the higher frequency of C-reactive protein in lepromatous leprosy patients, espicially, in the patients with reaction (Bush, 1958; Ross et al., 1959; Montestruc, 1960; Ross, 1964a, 1964b; Tamblyn and Hokama, 1969). In conjunction with these observations, a study of deter mination of serum immunglobulins, alpka2-macroglobulin and complement (C3) levels has been undertaken in 38 patients with lepromatous leprosy, 10 patients with tuberculoid leprosy and 20 normal healthy persons and in addition VDRL slide quantitative test and C-reactive protein test were performed in this investigation. Materials and method The study group consisted of 38 patients with lepromatous leprosy and 10 patients with tuberculoid leprosy who have been treated at the World Vision Special Skin Clinic, and 20 normal healthy persons as a control group. Determination of immunoglobulins (IgG, IgA, and IgM), alpha2-macroglobulin and complement (C3) levels; Single radial immunodiffusion method (Palmer and Woods, 1972) was applied for the measurement of IgG, IgM, alpha2-macroglobulin and complement (C3) levels, using the immunodiffusion plate (Hyland Lsb.). Briefly, each well was filled with the three reference sera and test sera to the level of the agar surface, and the plates were incubated for 16 to 18 hours in a moist chamber. After incubation, the diameter of the precipitin fing diameter of the three reference sera on 2-cycle semilogarithmic graph paper, and the concentration of test sera were determines by the reference curve. VDRL slide quantitative test; After heat-inactivation of sera at 56℃ for 30 min., 0.013ml of antigen (VDRL flocculation antigen, Sylvana Co.) was added to 0.04ml of 2-fold serially diluted serum and observed the highest dilution that produce a reactive after rotating 4 min. (Lennette er al., 1974). C-reactive protein test; C-reactive protein antiserum (Bacto C protein antiserum (Bacto C protein antisermm, Difco Lab.) was drawn up to the one third of 90mm long capillary tube (0.4mm inside diameter), and after wiping the tip clean equal amount of test serum was drawn up. The height of precipitate was measured after incubation at 4℃ overnight, followed incubation at 37℃ for 2 hours. Summary and conclusion The study of serum immunoglobulins, alpha2-macroglobulin and complement (C3) levels, as well as VDRL slide quantitative test and C-reactive protein test, was conducted with 38 patients with lepromatous leprosy, 10 patients with tuberculoid leprosy and 20 normal healthy persons in an attempt to define the hyperactive state of humoral immunity and the mechanism of lepra reaction in lepromatous leprosy, and the results are summarized as follows: 1. The serum IgG and IgA levels in lepromatous leprosy were higher significantly than those of the normal control. 2. No significant differences were observed on the levels of alpha2-macroglobulin between the lepromatous leprosy, tuberculoid leprosy and normal control groups, but the level was lower in patients with ENL than in the patients without ENL. 3. The serum level of complement (C3) in lepromatous leprosy was significantly higher than those of tuberculoid leprosy and normal control group, and a tendency of higher levels of complement (C3) was observable in lepromatous leprosy patients with ENL and higher bacterial index. 4. Of 38 lepromatous leprosy patients 10 (26.3%) were positive in C-reactive protein test. These 10 C-reactive protein positive cases included 5 (55.6%) cases out of 9 lepromatous leprosy without ENL. All cases of tuberculoid leprosy and normal control groups showed negative in C-reactive protein test. 5. Three (7.9%) cases out of 38 lepromatous leprosy patients were reactive in VDRL slide quantitative test, i.e., 1:32, 1:4, and 1:2 respectively.
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