天皰瘡抗體滴度與疾病活動(dòng)性及CD19、CD20的相關(guān)性研究
發(fā)布時(shí)間：2018-08-03 11:15
【摘要】： 天皰瘡是一組以天皰瘡抗體介導(dǎo)的器官特異性自身免疫性疾病,發(fā)病率雖然很低,但呈上升趨勢(shì)。好發(fā)于30-50歲中年人群,無明顯性別差異。臨床上主要分為四型：尋常型、增殖型、落葉型、紅斑型,但四型之間可相互轉(zhuǎn)化。此外,還有皰疹樣天皰瘡、IgA天皰瘡、藥物誘發(fā)的天皰瘡、副腫瘤性天皰瘡、流行性性天皰瘡(又名巴西天皰瘡)等亞型。該組疾病的共同特征有：臨床表現(xiàn)為皰壁薄、松弛易破的水皰或大皰形成；組織病理為棘層松解所致表皮內(nèi)水皰形成；直接免疫熒光檢查(indirect immunofluorescence,IIF)可見棘層細(xì)胞間IgG和C3沉積,取患者血清做間接免疫熒光檢查,可顯示血清中有抗表皮棘細(xì)胞間物質(zhì)抗體。 天皰瘡的發(fā)病機(jī)理尚未完全清楚。越來越多研究認(rèn)為在天皰瘡發(fā)病過程中,T細(xì)胞和B細(xì)胞相互作用、相互影響,產(chǎn)生針對(duì)自身細(xì)胞間物質(zhì)的抗體,從而導(dǎo)致疾病的發(fā)生。天皰瘡的抗原主要集中在橋粒,橋粒是角質(zhì)形成細(xì)胞間連接的主要結(jié)構(gòu),由相鄰細(xì)胞的細(xì)胞膜發(fā)生卵圓形致密增厚而共同構(gòu)成。由兩類蛋白質(zhì)構(gòu)成：一類是跨膜蛋白,位于橋粒芯(desmosomal core),主要由橋粒芯蛋白(desmoglein,Dsg)和橋粒芯膠蛋白(desmocollin,Dsc)構(gòu)成,它們形成橋粒的電子透明細(xì)胞間隙和細(xì)胞間接觸層；另一類為胞質(zhì)內(nèi)的橋粒斑(desmosomal plaque)蛋白,是盤狀附著板的組成部分,主要成分為橋粒斑蛋白(desmoplakin,DP)和橋粒斑珠蛋白(plakogloubin,PG)。天皰瘡抗體(Pemphigus antibody)與抗原結(jié)合后,通過細(xì)胞信號(hào)傳導(dǎo)途徑激活一系列蛋白水解酶,導(dǎo)致細(xì)胞間鏈接結(jié)構(gòu)水解,從而引起表皮棘層細(xì)胞相互分離、棘層松解及表皮內(nèi)水皰形成。鈣粘素是Ca2+依賴的細(xì)胞間連接分子,它對(duì)表皮細(xì)胞間粘附連接起重要作用。天皰瘡抗原的c DNA序列與鈣粘素有明顯的同源性,因此天皰瘡抗體也損害了表皮細(xì)胞間的粘連功能,導(dǎo)致棘層松解。棘層松解在PF患者較表淺,損害僅達(dá)表皮顆粒層。而在PV患者則深達(dá)表皮生發(fā)層。幾乎所有天皰瘡患者免疫病理均示角質(zhì)形成細(xì)胞間IgG和C3呈網(wǎng)狀沉積。 天皰瘡的診斷主要結(jié)合四個(gè)方面因素考慮：1、典型的臨床皮損表現(xiàn)；2、皮膚組織病理所示的棘層松解；3、免疫病理示角質(zhì)形成細(xì)胞間IgG和C3呈網(wǎng)狀沉積；4、組織上發(fā)現(xiàn)患者血清中或原位皮膚存在自身抗橋粒芯糖蛋白抗體。目前天皰瘡治療方法很多,但皮質(zhì)類固醇激素仍是目前的首選,開始劑量需較大,以期在1-2周內(nèi)抑制新的水皰形成,必要時(shí)可應(yīng)用皮質(zhì)類固醇激素沖擊療法或加用免疫抑制劑治療。根據(jù)患者皮損面積分為輕、中、重度并給與適合劑量的激素,以不出現(xiàn)新的水皰為宜。皮損面積體表面積10%為輕度,激素量為30mg/d；體表面積30%皮損面積體表面積50%為中度,激素量為60-80 mg/d；皮損面積體表面積50%為重度,激素量80 mg/d。此外,部分皮損面積很局限的天皰瘡患者在口服少量激素的同時(shí)加用激素軟膏或他克莫司軟膏外用可促進(jìn)皮損的愈合。 但目前天皰瘡診治中最大的問題是明確診斷和判斷疾病的活動(dòng)程度及預(yù)后。尼氏征對(duì)于天皰瘡的診斷具有較高的特異性,但其敏感性較差。直接免疫熒光(direct immunofluorescence,DIF)檢查是診斷天皰瘡的金標(biāo)準(zhǔn),但反復(fù)的皮膚活檢術(shù)給病人造成較大的痛苦,難以為廣大的患者所接受,也難以作為隨訪的標(biāo)準(zhǔn)。根據(jù)已知的天皰瘡發(fā)病機(jī)制,本實(shí)驗(yàn)嘗試檢測(cè)血循環(huán)中的天皰瘡抗體對(duì)天皰瘡患者進(jìn)行診斷,并探討其與疾病的活動(dòng)性的關(guān)系,以尋找出適合臨床診斷、判斷療效和隨訪的指標(biāo)。 目的： 1、通過酶聯(lián)免疫吸附(Enzyme-linked immunosorbent assay,ELISA)檢測(cè)天皰瘡患者血清中的抗體IgG滴度,了解其敏感性和特異性。 2、對(duì)天皰瘡患者的病情活動(dòng)度進(jìn)行評(píng)分,探討其與天皰瘡抗體滴度的相關(guān)性。 3、通過流式細(xì)胞儀(Flow cytometry,FCM)檢測(cè)天皰瘡患者血清中CD19、CD20的表達(dá)水平,探討天皰瘡抗體滴度與CD19、CD20表達(dá)水平的相關(guān)性。 方法： 1、收集2009-1至2010-2之間門診及住院并確診的38例天皰瘡患者及非天皰瘡患者22人的血清,離心后用天皰瘡抗體檢測(cè)試劑盒(MBL公司),通過ELISA法檢測(cè)60份血清中天皰瘡抗體IgG的滴度。了解該方法與臨床的相符性,并探討其敏感性和特異性。 2、對(duì)確診的天皰瘡患者進(jìn)行病情活動(dòng)度評(píng)分,分析疾病病情活動(dòng)評(píng)分與天皰瘡抗體滴度進(jìn)行相關(guān)性。 3、將38例天皰瘡患者中的28位患者的血清經(jīng)流式細(xì)胞技術(shù)檢測(cè)其中CD19、CD20的表達(dá)水平,并與之前檢測(cè)的相應(yīng)的天皰瘡抗體滴度進(jìn)行多元相關(guān)性分析。 結(jié)果： 1、診斷的符合度、敏感性和特異性：38例天皰瘡患者中有37例陽性,1例陰性。22例非天皰瘡患者則均為陰性。該方法的診斷符合率為98.3%,敏感性為97.4%,特異性為100%。 2、病情活動(dòng)度評(píng)分與天皰瘡抗體滴度的Pearson雙變量相關(guān)性分析：病情活動(dòng)度評(píng)分與天皰瘡抗體滴度之間的相關(guān)性具有統(tǒng)計(jì)學(xué)意義(P0.001),相關(guān)系數(shù)r=0.671,直線回歸方程為：病情活動(dòng)評(píng)分=1.079+0.671×抗體滴度。 3、天皰瘡抗體滴度與血清CD19、CD20表達(dá)水平的多元回歸分析：天皰瘡抗體滴度與血清CD19、CD20表達(dá)水平的的相關(guān)性亦具有統(tǒng)計(jì)學(xué)意義(P0.05),將數(shù)據(jù)進(jìn)行對(duì)數(shù)轉(zhuǎn)換后,其復(fù)相關(guān)系數(shù)R=0.55,R2=0.302,lgCD19(x1)的偏回歸系數(shù)為0.518,lgCD20(x2)的偏回歸系數(shù)為0.066,截距為1.674,得出回歸方程為：Lg抗體滴度=1.674+0.518×LgCD19+0.066×LgCD20。 結(jié)論： 1、該實(shí)驗(yàn)進(jìn)一步證實(shí)了天皰瘡抗體用于天皰瘡的診斷具有較高的敏感性及特異性,與臨床符合率較高,對(duì)疾病的診斷具有重要的意義。 2、天皰瘡抗體抗體滴度越高,疾病的活動(dòng)性越強(qiáng),與疾病活動(dòng)度相平行,具有較高的相關(guān)性�？捎糜诩膊』顒�(dòng)度的判斷、治療效果的評(píng)價(jià)隨訪的指標(biāo)。 3、天皰瘡抗體滴度與患者血清中的CD19、CD20的表達(dá)水平亦具有相關(guān)性,CD19、CD20的表達(dá)水平越高,血清中的抗體的滴度也越高。
[Abstract]:Pemphigus is an organ specific autoimmune disease mediated by pemphigus antibody. Although the incidence of pemphigus is very low, the incidence of pemphigus is on the rise. There are no significant gender differences in the 30-50 year old middle-aged people. It is mainly divided into four types: vulgaris, proliferating, deciduous, erythema, but the type four can be transformed from each other. In addition, there are herpes samples. Pemphigus, pemphigus IgA, pemphigus induced by drugs, paraneoplastic pemphigus, and pemphigus epidemic (also known as pemphigus Brazil). The common characteristics of the disease are: the clinical manifestation is the thin wall of the blister, the loosening blister or bulla; the histopathology is the formation of the blister caused by the spinous release of the epidermis; direct immunofluorescence. Indirect immunofluorescence (IIF) shows the deposition of IgG and C3 between the spinous cells, and the indirect immunofluorescence examination of the patient's serum shows that there are anti epidermal acanthosis antibodies in the serum.
The pathogenesis of pemphigus is not completely clear. More and more studies have shown that in the process of pemphigus, T and B cells interact and interact with each other to produce antibodies against their own intercellular substances, which lead to the occurrence of disease. The antigen of pemphigus is mainly concentrated in the pemphigus, and the pemphigus is the main junction of the keratinocytes. The structure, composed of two kinds of proteins, consists of two kinds of proteins: a kind of transmembrane protein, located in the grain core (desmosomal core), mainly composed of desmoglein, Dsg and desmocollin, Dsc, which form the transparent space and intercellular space of the bridging particles. The contact layer; the other kind is the desmosomal plaque protein in the cytoplasm, which is a component of the disc attachment plate. The main components are desmoplakin, DP and plakogloubin, PG. Pemphigus antibody (Pemphigus antibody) activates a series of protein water through the cell signal transduction pathway after the combination of the antibody (Pemphigus antibody) with the anti original. The hydrolytic enzyme leads to the hydrolysis of intercellular links, resulting in the isolation of the epidermal acanthosis cells, the acanthosis and the formation of the blisters in the epidermis. The cadherin is a Ca2+ dependent intercellular junction molecule, which plays an important role in the adhesion and connection between the epidermis cells. The C DNA sequence of pemphigus antigen has obvious homology with the cadherin, so pemphigus is resistant to pemphigus. The body also damaged the adhesion function between the epidermal cells and resulted in the acanthosis. The acanthosis was superficial in the PF patients and the damage was only to the epidermis. In the patients with PV, the epidermis was deep. The immunhistopathology of almost all pemphigus showed that the IgG and C3 were meshed between the keratinocytes.
The diagnosis of pemphigus was mainly combined with four factors: 1, typical clinical skin lesions, 2, acanthosis of the skin histopathology; 3, IgG and C3 in the keratinocytes of the immunhistopathology showed reticular deposition; 4, the tissue was found in the sera or in situ in its own anti pemphigoprotein antibody. There are many treatments, but corticosteroids are still the first choice. The initial dose should be large in order to inhibit the formation of new blisters within 1-2 weeks. It is necessary to use corticosteroid shock therapy or immunosuppressive therapy when necessary. According to the area of skin lesions, the patients are divided into light, medium, severe and given the suitable dose of hormone, in order not to appear. The surface area of the skin area was mild, the surface area of the skin lesion was 10%, the volume of the body surface area was 30mg/d, the body surface area 30% skin area 50% was moderate, the hormone amount was 60-80 mg/d, the skin area surface area 50% was 50%, the hormone was 80 mg/d., and the patients with the limited area of skin lesions were combined with a small amount of hormone at the same time. External application of plain ointment or Tacrolimus Ointment can promote the healing of skin lesions.
But at present, the biggest problem in the diagnosis and treatment of pemphigus is to diagnose and judge the degree of activity and prognosis of the disease. Nil's sign has a high specificity for the diagnosis of pemphigus, but its sensitivity is poor. Direct immunofluorescence (direct immunofluorescence, DIF) is the gold standard for the diagnosis of pemphigus, but repeated skin biopsy is given to the patients. According to the known pathogenesis of pemphigus, this experiment is trying to detect pemphigus antibodies in the blood circulation to diagnose pemphigus, and to explore the relationship between pemphigus and the activity of the disease in order to find a suitable clinical diagnosis, to judge the curative effect and follow it. The target of the visit.
Objective:
1. Enzyme-linked immunosorbent assay (ELISA) was used to detect the antibody IgG titer in the serum of patients with pemphigus.
2, we assessed the degree of disease activity in pemphigus patients and explored the correlation between pemphigus antibody titer and pemphigus antibody titer.
3, the expression level of CD19 and CD20 in the serum of pemphigus patients was detected by Flow cytometry (FCM), and the correlation between the tipper titer antibody titer and the expression of CD19 and CD20 was investigated.
Method:
1, the serum of 38 pemphigus patients and 22 pemphigus patients were collected from 2009-1 to 2010-2 in outpatients and hospitalized. After centrifugation, the titer antibody test kit (MBL company) was used to detect the titer of the pemphigus antibody IgG in 60 sera and to understand the coincidence of this method with the clinical and its sensitivity and specificity.
2. The patients with pemphigus were assessed by disease activity score, and the correlation between disease activity score and pemphigus antibody titer was analyzed.
3, the serum level of CD19 and CD20 was detected by serum flow cytometry in 28 patients with pemphigus, and the correlation was analyzed with the correlation of the antibody titer of pemphigus which was detected before.
Result:
1, diagnostic accuracy, sensitivity and specificity: 37 cases were positive in 38 pemphigus patients and 1 negative.22 cases were negative. The diagnostic coincidence rate of this method was 98.3%, sensitivity was 97.4%, and the specificity was 100%..
2, the correlation of Pearson bivariate analysis of disease activity score and pemphigus antibody titer: the correlation between disease activity score and pemphigus antibody titer was statistically significant (P0.001), the correlation coefficient was r=0.671, and the linear regression equation was =1.079 +0.671 x antibody titer.
3, multiple regression analysis of the titer's titer antibody titer and the level of serum CD19, CD20 expression: the correlation between the tipper titer antibody titer and the serum CD19, CD20 expression level also has statistical significance (P0.05). After the logarithmic conversion of the data, the partial regression coefficient of the correlation coefficient R=0.55, R2=0.302, lgCD19 (x1) is 0.518, lgCD20 (x2) partial regression The coefficient is 0.066 and the intercept is 1.674. The regression equation is: Lg antibody titer =1.674+0.518 * LgCD19+0.066 * LgCD20.
Conclusion:
1, the experiment further confirmed that pemphigus antibody has high sensitivity and specificity in the diagnosis of pemphigus, and it has a higher coincidence with the clinic. It is of great significance for the diagnosis of the disease.
2, the higher the titer antibody titer, the stronger the activity of the disease, and the higher correlation with the degree of disease activity. It can be used to judge the degree of disease activity and evaluate the outcome of the treatment.
3, the titer of pemphigus was also associated with the expression level of CD19 and CD20 in the patient's serum. The higher the expression level of CD19, the higher the level of CD20, the higher the titer of the antibody in the serum.
【學(xué)位授予單位】：南方醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2010
【分類號(hào)】：R758.66
本文編號(hào)：2161573
[Abstract]:Pemphigus is an organ specific autoimmune disease mediated by pemphigus antibody. Although the incidence of pemphigus is very low, the incidence of pemphigus is on the rise. There are no significant gender differences in the 30-50 year old middle-aged people. It is mainly divided into four types: vulgaris, proliferating, deciduous, erythema, but the type four can be transformed from each other. In addition, there are herpes samples. Pemphigus, pemphigus IgA, pemphigus induced by drugs, paraneoplastic pemphigus, and pemphigus epidemic (also known as pemphigus Brazil). The common characteristics of the disease are: the clinical manifestation is the thin wall of the blister, the loosening blister or bulla; the histopathology is the formation of the blister caused by the spinous release of the epidermis; direct immunofluorescence. Indirect immunofluorescence (IIF) shows the deposition of IgG and C3 between the spinous cells, and the indirect immunofluorescence examination of the patient's serum shows that there are anti epidermal acanthosis antibodies in the serum.
The pathogenesis of pemphigus is not completely clear. More and more studies have shown that in the process of pemphigus, T and B cells interact and interact with each other to produce antibodies against their own intercellular substances, which lead to the occurrence of disease. The antigen of pemphigus is mainly concentrated in the pemphigus, and the pemphigus is the main junction of the keratinocytes. The structure, composed of two kinds of proteins, consists of two kinds of proteins: a kind of transmembrane protein, located in the grain core (desmosomal core), mainly composed of desmoglein, Dsg and desmocollin, Dsc, which form the transparent space and intercellular space of the bridging particles. The contact layer; the other kind is the desmosomal plaque protein in the cytoplasm, which is a component of the disc attachment plate. The main components are desmoplakin, DP and plakogloubin, PG. Pemphigus antibody (Pemphigus antibody) activates a series of protein water through the cell signal transduction pathway after the combination of the antibody (Pemphigus antibody) with the anti original. The hydrolytic enzyme leads to the hydrolysis of intercellular links, resulting in the isolation of the epidermal acanthosis cells, the acanthosis and the formation of the blisters in the epidermis. The cadherin is a Ca2+ dependent intercellular junction molecule, which plays an important role in the adhesion and connection between the epidermis cells. The C DNA sequence of pemphigus antigen has obvious homology with the cadherin, so pemphigus is resistant to pemphigus. The body also damaged the adhesion function between the epidermal cells and resulted in the acanthosis. The acanthosis was superficial in the PF patients and the damage was only to the epidermis. In the patients with PV, the epidermis was deep. The immunhistopathology of almost all pemphigus showed that the IgG and C3 were meshed between the keratinocytes.
The diagnosis of pemphigus was mainly combined with four factors: 1, typical clinical skin lesions, 2, acanthosis of the skin histopathology; 3, IgG and C3 in the keratinocytes of the immunhistopathology showed reticular deposition; 4, the tissue was found in the sera or in situ in its own anti pemphigoprotein antibody. There are many treatments, but corticosteroids are still the first choice. The initial dose should be large in order to inhibit the formation of new blisters within 1-2 weeks. It is necessary to use corticosteroid shock therapy or immunosuppressive therapy when necessary. According to the area of skin lesions, the patients are divided into light, medium, severe and given the suitable dose of hormone, in order not to appear. The surface area of the skin area was mild, the surface area of the skin lesion was 10%, the volume of the body surface area was 30mg/d, the body surface area 30% skin area 50% was moderate, the hormone amount was 60-80 mg/d, the skin area surface area 50% was 50%, the hormone was 80 mg/d., and the patients with the limited area of skin lesions were combined with a small amount of hormone at the same time. External application of plain ointment or Tacrolimus Ointment can promote the healing of skin lesions.
But at present, the biggest problem in the diagnosis and treatment of pemphigus is to diagnose and judge the degree of activity and prognosis of the disease. Nil's sign has a high specificity for the diagnosis of pemphigus, but its sensitivity is poor. Direct immunofluorescence (direct immunofluorescence, DIF) is the gold standard for the diagnosis of pemphigus, but repeated skin biopsy is given to the patients. According to the known pathogenesis of pemphigus, this experiment is trying to detect pemphigus antibodies in the blood circulation to diagnose pemphigus, and to explore the relationship between pemphigus and the activity of the disease in order to find a suitable clinical diagnosis, to judge the curative effect and follow it. The target of the visit.
Objective:
1. Enzyme-linked immunosorbent assay (ELISA) was used to detect the antibody IgG titer in the serum of patients with pemphigus.
2, we assessed the degree of disease activity in pemphigus patients and explored the correlation between pemphigus antibody titer and pemphigus antibody titer.
3, the expression level of CD19 and CD20 in the serum of pemphigus patients was detected by Flow cytometry (FCM), and the correlation between the tipper titer antibody titer and the expression of CD19 and CD20 was investigated.
Method:
1, the serum of 38 pemphigus patients and 22 pemphigus patients were collected from 2009-1 to 2010-2 in outpatients and hospitalized. After centrifugation, the titer antibody test kit (MBL company) was used to detect the titer of the pemphigus antibody IgG in 60 sera and to understand the coincidence of this method with the clinical and its sensitivity and specificity.
2. The patients with pemphigus were assessed by disease activity score, and the correlation between disease activity score and pemphigus antibody titer was analyzed.
3, the serum level of CD19 and CD20 was detected by serum flow cytometry in 28 patients with pemphigus, and the correlation was analyzed with the correlation of the antibody titer of pemphigus which was detected before.
Result:
1, diagnostic accuracy, sensitivity and specificity: 37 cases were positive in 38 pemphigus patients and 1 negative.22 cases were negative. The diagnostic coincidence rate of this method was 98.3%, sensitivity was 97.4%, and the specificity was 100%..
2, the correlation of Pearson bivariate analysis of disease activity score and pemphigus antibody titer: the correlation between disease activity score and pemphigus antibody titer was statistically significant (P0.001), the correlation coefficient was r=0.671, and the linear regression equation was =1.079 +0.671 x antibody titer.
3, multiple regression analysis of the titer's titer antibody titer and the level of serum CD19, CD20 expression: the correlation between the tipper titer antibody titer and the serum CD19, CD20 expression level also has statistical significance (P0.05). After the logarithmic conversion of the data, the partial regression coefficient of the correlation coefficient R=0.55, R2=0.302, lgCD19 (x1) is 0.518, lgCD20 (x2) partial regression The coefficient is 0.066 and the intercept is 1.674. The regression equation is: Lg antibody titer =1.674+0.518 * LgCD19+0.066 * LgCD20.
Conclusion:
1, the experiment further confirmed that pemphigus antibody has high sensitivity and specificity in the diagnosis of pemphigus, and it has a higher coincidence with the clinic. It is of great significance for the diagnosis of the disease.
2, the higher the titer antibody titer, the stronger the activity of the disease, and the higher correlation with the degree of disease activity. It can be used to judge the degree of disease activity and evaluate the outcome of the treatment.
3, the titer of pemphigus was also associated with the expression level of CD19 and CD20 in the patient's serum. The higher the expression level of CD19, the higher the level of CD20, the higher the titer of the antibody in the serum.
【學(xué)位授予單位】：南方醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2010
【分類號(hào)】：R758.66
【參考文獻(xiàn)】
相關(guān)期刊論文 前3條
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2 李渝,刁慶春,王魯,鐘白玉,唐書謙,葉慶佾,劉榮卿;天皰瘡抗體亞類在天皰瘡發(fā)病中的作用[J];中華皮膚科雜志;2000年04期
3 聶祝湘,劉榮卿,葉慶佾,唐書謙,鐘白玉;天皰瘡抗體對(duì)橋粒影響的動(dòng)態(tài)觀察[J];中華醫(yī)學(xué)雜志;1996年09期
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