Natural antibodies in sera of patients with systemic lupus erythematosus

Sera obtained from fifty-five patients with active systemic lupus erythematosus (SLE) and from four patients with mixed connective tissue disease (MCTD) previously shown by immunofluorescence and by double immunodiffusion to possess antinuclear antibodies, were tested for the presence of natural antibodies of IgG, IgA, and IgM isotypes. Antibody activity to actin, myosin, DNA, TNP, albumin, and tubulin was examined, using an enzyme-linked immunosorbent assay (ELISA). It was found that, in comparison with the antibody titers in normal sera, most of the SLE and MCTD sera possessed statistically greater amounts of IgG, IgA, and IgM antibodies directed against all the antigens tested. Furthermore, the IgG, IgA, and IgM antibody activity to DNA and TNP, compared to that found with all the other antigens, was significantly higher. Antibodies reacting with a saline extract of calf thymus (ECT) were studied by ELISA and by immunodiffusion. No correlation was observed between the natural antibody titers and the serum antibody levels to ECT detected either by ELISA or by immunodiffusion.     

Utilization of hantavirus antibody results generated over a five-year period to develop an improved serologic algorithm for detecting acute Sin Nombre hantavirus infection

In the United States most cases of hantavirus pulmonary syndrome (HPS) are caused by the Sin Nombre virus (SNV) and are typically identified by serology. The goal of this study was to assess the performance of our hantavirus serologic testing algorithm by reviewing results generated over five years. Sera were screened for pan-hantavirus immunoglobulin (Ig)G and IgM by enzyme-linked immunosorbent assay (ELISA). Screen IgG+ sera were then tested by immunoblot for SNV glycoprotein-specific IgG, and screen IgM+ sera were tested for SNV-specific IgM using an ELISA that measured differential reactivity to SNV and Seoul nucleocapsid proteins. Although only 13% of sera were positive in one or both screening assays, 85% of screen+sera lacked SNV antibodies. Nearly all (97%) screen IgM-IgG+ samples lacked SNV IgG, and 90% of screen IgM+IgG- samples lacked SNV IgM. However, SNV IgM testing of screen IgM+IgG- samples appears to be necessary, since this test identified nine of 37 patients with acute HPS (based on clinical feedback). A screen IgM+IgG+ result was a good predictor of SNV antibody detection and acute HPS. These findings were used to design a modified algorithm that identified all 37 patients with acute HPS, but reduced the number of specimens that required SNV antibody testing by 42%.     

Natural autoantibodies against Tamm-Horsfall glycoprotein in normal individuals in relation to age and in adult patients with kidney diseases

IgM and IgG natural antibodies to Tamm-Horsfall glycoprotein (THP) were found in serum samples of all healthy individuals tested by the ELISA technique. The IgM anti-THP antibody level was higher in the group 1-20 years old than the IgG anti-THP. The IgG anti-THP rose with increase in age (greater than 21 years old groups) and then the IgG and IgM anti-THP activity over aging remained constant. The natural anti-THP antibodies possess a lower degree of specificity and/or avidity than induced antibodies. The antibody titers against THP determined in 61 adult patients with chronic kidney diseases was significantly lower than that in adult controls. This low level of naturally occurring THP antibodies appears to be a general phenomenon. In these patients, diminished antibody levels appeared against a panel of self (collagen, fibronectin, THP) and non self (bovine gamma globulin (BGG), ovalbumin (OVA)) antigens as compared with normal controls. The low levels of these antibodies are not associated with a concomitant drop of IgG and IgM in their sera.     

Direct evidence that decreased serum opsonization of Streptococcus pneumoniae via the alternative complement pathway in sickle cell disease is related to antibody deficiency.

Two approaches were used to demonstrate that reduction in serum opsonization of Streptococcus pneumoniae via the alternative complement pathway in children with sickle cell disease is related to a deficiency of antibodies to pneumococcal capsular polysaccharide. First, opsonization of S. pneumoniae mediated by the alternative pathway in patients' sera was restored to normal by addition of the purified IgG or IgM fraction of goat antiserum to capsular polysaccharide of the homologous serotype. Secondly, IgG antibody titers to capsular polysaccharide in patients' sera correlated significantly with alternative pathway-mediated opsonization; the correlation between titers of IgM anticapsular antibodies and opsonization approached statistical significance. The sum of the IgG and IgM anticapsular antibody titers correlated most significantly with opsonization. Our results suggest that reduction in alternative pathway-mediated opsonization in sera from children with sickle cell disease is related to low levels of both IgG and IgM anticapsular antibodies.     

肺炎嗜衣原体蛋白酶样活性因子重组蛋白的血清学诊断价值初探

目的克隆、表达肺炎嗜衣原体（Cpn）蛋白酶样活性因子（CPAF）的免疫优势区基因（CPAFm），初步评价其在血清学诊断中的应用价值。方法构建pGEX6p-2／CPAFm重组质粒，诱导表达重组蛋白，十二烷基磺酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）鉴定结果；将纯化蛋白免疫新西兰兔，间接酶联免疫吸附试验（ELISA）检测其免疫原性；与人抗Cpn抗血清反应分析其免疫反应性。间接ELISA法检测Cpn IgG和沙眼衣原体（Ct）阳性血清。结果高效表达和纯化出一相对分子质量约为51.3×10^3的重组蛋白，蛋白质印迹法（Western blot）证明其能与人抗Cpn IgC抗体发生反应；在被免疫的新西兰兔体内，特异性IgG抗体的滴度为1：16000以上，间接ELISA法检测40例Cpn IgG参考血清，阴性和阳性结果的符合率均为100．0%（20／20）；与微量免疫荧光法（MIF）对照，检测300例临床血清标本中的IgG抗体，符合率为98．3％；检测Ct阳性血清未见交叉反应。结论表达的Cpn CPAF免疫优势区重组蛋白具有良好的免疫活性，在Cpn的血清学诊断中具有较高的应用价值。     

Antibiodies of the IgG, IgM, and IgA classes in newborn and adult sera reactive with gram-negative bacteria

Umbilical cord serum and adult serum antibodies reactive with heat-stable somatic antigens of Gram-negative bacteria (Neisseria gonorrhoeae, Escherichia coli, and Salmonella typhosa) were assayed by using an indirect fluorescent antibody test. Reactive IgG, IgM, and IgA antibodies were identified by using fluoresceinconjugated antisera specific for these immunoglobulin classes.IgG antibody titers in cord serum approximated those found in the corresponding maternal sera. IgM and IgA antibodies were present in adult sera but were not demonstrable or were present only in small amounts in cord sera. The presence of IgG and IgM antibodies reactive with Gram-negative bacteria was confirmed by the testing of purified 7S and 19S fractions. In addition, both IgG and IgM reactivities were inhibited by the prior incubation of serum with purified specific lipopolysaccharide preparations.The ubiquity and magnitude of these natural IgG antibodies in the sera of both adults and neonates have apparently eluded detection in previous studies. The use of bactericidal and agglutination tests, which are apparently more sensitive to the presence of IgM than to IgG antibodies, may account for the failure of previous studies to detect adult and cord IgG antibodies reactive with somatic antigens of Gram-negative bacteria. The presence of these IgG antibodies may be correlated with the resistance to infection demonstrated by most newborns as they are challenged by the septic extrauterine environment.     

非典型肺炎血清SARS冠状病毒抗体的检测及其意义

【目的】探讨非典型肺炎患者血清SARS冠状病毒抗体IgM、IgG检测意义。【方法】应用ELISA法对16例SARS患者、147例发热隔离患者、23例体格检查健康者进行血清SARS抗体IgM、IgG检测。【结果】12例SARS患者入院时8例检出SARS抗体IgM阳性,出院时9例检出SARS抗体IgG阳性;其中7例SARS患者康复出院半年后复查均检测出高滴度SARS抗体IgG,同时有2例仍检出SARS抗体IgM。2例重症患者血清动态结果观察到,从入院到出院的标本均同时检测到高滴度的SARS IgM、IgG;2例轻症SARS患者各只检测到一次SARS抗体IgM弱阳性;发热隔离区患者有6例从入院至出院均检测到SARS抗体IgG,其中两例滴度较高。健康体检者无一例SARS抗体阳性。【结论】ELISA检测SARS冠状病毒抗体IgM、IgG可作为一种辅助鉴定非典型肺炎患者的临床检测和确诊手段。     

Analysis of cross-reactivity among flaviviruses using sera of patients with dengue showed the importance of neutralization tests with paired serum samples for the correct interpretations of serological test results for dengue

Dengue is a febrile illness caused by the dengue virus (DENV) that belongs to the genus Flavivirus in the family Flaviviridae. Cross-reactivity between flaviviruses poses a challenge while interpreting serological test results. In the present study, the cross-reactivity of sera of the patients with dengue, who traveled from Japan to DENV-endemic countries, was analyzed by using an enzyme-linked immunosorbent assay (ELISA) and neutralization test (NT). Sixteen serum samples were collected from patients with dengue and were tested for: i) IgM antibodies against Zika virus (ZIKV), West Nile virus (WNV), Japanese encephalitis virus (JEV), and tick-borne encephalitis virus (TBEV) using IgM ELISA, ii) IgG antibody against TBEV using IgG ELISA, and iii) neutralizing antibody against ZIKV, WNV, TBEV, and JEV. Among the 16 samples tested using ELISA, seven samples were IgM-positive for at least one of the other flaviviruses, and nine samples were IgG-positive for TBEV. Neutralizing antibody titers (NATs) against ZIKV, WNV, and TBEV were one-fourth or lower than those against the causative DENV in all samples. The NATs against JEV were one-fourth or lower than those against the causative DENV in six convalescent-phase serum sample among the seven convalescent-phase serum samples. The NAT against DENV of the residual one convalescent-phase serum was similar to that against JEV and that against JEV of its relevant acute-phase serum sample. These results showed that NTs with paired serum samples are important to correctly interpret the serological test results for DENV.     

Anti chlamydia antibodies in patients with thoracic and abdominal aortic aneurysms

BACKGROUND: Chlamydia species are suspected of being involved in the pathogenesis and progression of aortic aneurysms. We investigated serum levels of Chlamydia antibodies in patients with thoracic aortic aneurysms (TAA) and abdominal aortic aneurysms (AAA) compared to levels in healthy individuals. METHODS: We included 35 consecutive patients with TAA, 42 patients with AAA and 42 age- and sex-matched healthy controls in a case control study. Serum antibodies (IgM and IgG) against Chlamydia lipopolysaccharide (LPS), Chlamydia pneumoniae and Chlamydia trachomatis were measured by recombinant ELISA and quantified by measurement of optical density. RESULTS: Patients with TAA exhibited median immunoglobulin levels against Chlamydia LPS (IgM 0.090, IgG 0.266), C. pneumoniae (IgM 0.023, IgG 0.264) and C. trachomatis (IgG 0.247) comparable to those of healthy subjects [Chlamydia LPS IgM 0.209 (p = 0.1), IgG 0.301 (p = 0.2); C. pneumoniae IgM 0.051 (p = 0.07), IgG 0.516 (p = 0.1); C. trachomatis IgG 0.153 (p = 0.2)]. Patients with AAA had higher serum levels of IgG against Chlamydia LPS (0.560) compared to healthy individuals [0.301 (p = 0.04)], but no significant elevation of antibodies against C. pneumoniae [IgM 0.029 (p = 0.1), IgG 0.545 (p = 0.9)] and C. trachomatis [IgG 0.219 (p = 0.3)]. CONCLUSION: Thoracic aortic aneurysms were not associated with signs of Chlamydia infection or immunopathogenicity. In contrast, patients with abdominal aortic aneurysms exhibited elevated levels of immunoglobulin against Chlamydia LPS, reflecting an unspecific Chlamydia immunopathogenicity. However, elevated levels of antibodies against distinct Chlamydia species were also not found in AAA patients.     

Hepatitis E antibody profiles in serum and urine

The aim of this study was to evaluate anti-HEV antibody profiles in urine specimens in comparison to corresponding serum samples to assess the utility of urine as a clinical specimen. Paired serum and urine specimens from 71 hepatitis E patients, 33 non-E hepatitis patients, 63 patients with nonhepatic diseases, and 26 healthy individuals were tested by recombinant HEV protein (55 kD)-based indirect enzyme-linked immunosorbent assay (ELISA). Uronegativity for anti-HEV IgM was noted in 71 (100%) serologically confirmed patients with hepatitis E. Hepatitis E patients (10/10) showed urinary absence or very low levels of total IgM by capture ELISA, suggesting absence or low levels of filtration, and/or local synthesis, and/or transudation of IgM in urine during infection. When these patients were tested for total IgG and IgA, microquantities of immunoglobulins were noted in all urine samples (10/10 for each). However, the proportions of uropositivity for anti-HEV IgG and IgA in hepatitis E patients were low and indicated only 21.42% and 49.33% concordance with seropositivity, respectively. Control groups also showed low and variable uropositivity for anti-HEV IgG and IgA. Overall, HEV-specific antibodies exhibited by serum in recent and past infections were not found in urine. The study demonstrated the inadequacy of urine specimens for detection of hepatitis E antibodies.     

Serological response to plasmid-encoded antigens in children and adults with shigellosis

The immunological response to plasmid-encoded antigens of virulent Shigella was determined in Thai children less than 4 yr of age and in Thai adults by immunoblot analysis and ELISA. Forty-two percent (8/19) of Thai children and 4% (1/22) of Thai adults with shigellosis developed a greater than or equal to 4-fold rise in IgG antibody titer to water-extracted antigens of Shigella flexneri M90T by ELISA (p = 0.006). Two children and one lactating mother with shigellosis developed a 4-fold rise in serum IgA antibody titers to water-extracted antigens of M90T. The results of the ELISA were confirmed by immunoblot analysis in all of the 41 paired sera examined. Five patients developed IgA, and four developed IgM, antibodies as detected by immunoblot analysis, that were not detected by ELISA. The reciprocal log2 geometric mean titers of antibodies to plasmid-encoded antigens in acute sera was higher in Thai adults than Thai children: IgG 7,265 versus 1,659; IgM 879 versus 480; and IgA 662 versus 60 (p less than 0.001). Thai adults had high titers of antibodies to plasmid-encoded antigens in their acute sera, but were susceptible to Shigella infections, although they were historically less susceptible than Thai children.     

广州地区献血人群人类细小病毒B19感染情况及病毒载量分析

目的了解广州地区献血者HPVB19感染状况。方法采用EIA对献血者血进行HPVB19 IgG、IgM筛查,并用PCR法对HPV B19抗体阳性血样进行HPV B19 DNA检测。结果HPV B19 IgG阳性率为38.6%(679/1760),HPVB19 IgM阳性率为1.9%(33/1760),两者有显著差异(P<0.001)。33例HPVB19 IgM阳性样本,HPVB19 DNA阳性检出率为63.6%(21/33);56例HPVB19 IgG阳性样本,HPVB19 DNA阳性检出率为1.8%(1/56),两者有显著差异(P<0.001)。21例HPV B19 DNA阳性样本中,病毒载量≥1×104Copies/ml占51.9%(13/21)。结论广州地区献血者HPV B19病毒既往感染率较高,但HPV B19病毒急慢性感染率较低。     

肺炎嗜衣原体三种不同抗原的临床诊断价值评价

目的 研究肺炎嗜衣原体3种重组抗原Cpn0146、Cpn0147及Cpn0308应用于肺炎嗜衣原体感染血清学诊断中的价值.方法 构建pGEX6p-2/Cpn0146、Cpn0147、Cpn0308重组质粒,并诱导表达重组蛋白,采用间接ELISA法和免疫印迹法(Western-blot)分析其免疫原性及其免疫反应性.建立分别以3种重组蛋白为包被抗原的间接ELISA法,与肺炎嗜衣原体IgG商品诊断试剂盒平行检测183份呼吸道感染患者的血清标本及32份沙眼衣原阳性患者血清标本,比较各方法检出准确性及阳性率.结果 GST-Cpn0146、Cpn0147、Cpn0308免疫兔血清中特异性IgG抗体效价滴度分别达1:6 400、1:12 800和1:12 800以上;以3种重组蛋白为包被抗原的ELISA法准确性分别为92.3%、94.5%、96.7%.各方法检测出阳性率结果显示,Cpn0146组的阳性率为38.8%(71/183)、与肺炎嗜衣原体IgG试剂盒的阳性率比较差异有统计学意义(x~2=12.07,P<0.05);Cpn0147阳性率40.9%(75/183),与肺炎嗜衣原体IgG试剂盒的阳性率比较差异有统计学意义(x~2=8.10,P<0.05);Cpn0308阳性率44.8%(75/183),与肺炎嗜衣原体IgG试剂盒的阳性率比较差异无统计学意义(x~2=0.57,P>0.05).结论 Cpn0308在肺炎嗜衣原体感染血清学诊断实验中表现出良好的准确性(96.2%)和较高的阳性率(44.8%),可望用于进一步研制肺炎嗜衣原体诊断试剂盒.     

Patterns of anti-phospholipid antibody specificities.

To determine the importance of assaying for anti-phospholipid (PL) antibodies other than anti-cardiolipin antibodies in patients with putative anti-phospholipid antibody syndrome (APA), 1,513 serum samples from over 399 patients with the potential for thromboembolic disease were tested for IgM, IgG or IgA class antibodies to the following phospholipids: cardiolipin, phosphatidylserine, phosphatidylinositol, phosphatidylethanolamine, phosphatidylglycerol and phosphatidic acid. Non-specific binding was subtracted from total binding of antibodies to each PL to eliminate false positive ELISA results. Only patients whose sera contained antibody levels which were greater than three standard deviations above the means were considered positive. Patients who were positive for anti-PL antibody had antibodies to at least one of six phospholipids tested. Approximately 60% of the positive samples contained antibodies to a phospholipid other than cardiolipin. For IgG class, antibodies to phosphatidylserine were the most prevalent followed by antibodies to phosphatidylethanolamine and cardiolipin. These findings indicate that anti-phospholipid antibody analysis should be performed for more than anti-cardiolipin antibody to detect patients with APA.     

Evaluation of commercial antiglobulin sera over a two-year period. Part II. Anti-IgG and anti-IgM levels and undesirable contaminating antibodies

Antiglobulin sera, from nine different manufacturers, have been tested over a two-year period for their ability to detect different nonagglutinating, IgG and IgM blood group antibodies and for the presence of undesirable antibodies that cause the agglutination of nonglobulin coated red blood cells. There are not so many differences in anti-IgG levels among the sera as there are differences in anticomplement levels. Over the two-year period, there have been, in general, increases in the amounts of anti-IgM antibodies in the sera tested. Several of the sera, apparently prepared by dilution of raw rabbit serum and not by adsorption, contain antibodies that cause the agglutination of red blood cells not coated with IgG, IgM, or any of the components of complement.     

Evaluation of newly developed ELISA using "MESACUP-2 test mitochondrial M2" kit for the diagnosis of primary biliary cirrhosis

OBJECTIVES: An enzyme-linked immunosorbent assay (ELISA) using MESACUP-2 Test Mitochondria M2 kit (new-M2 ELISA) has recently become commercially available. The aim of this study was to evaluate the clinical utility of this newly developed ELISA for the diagnosis of primary biliary cirrhosis (PBC). DESIGN AND METHODS: We tested the immunoreactivity of sera from 82 Japanese PBC patients to the 2-oxo-acid dehydrogenase complex (2-OADC) enzymes by indirect immunofluorescence, enzyme inhibition assay using commercially available TRACE Enzymatic Mitochondrial Antibody (M2) Assay (EMA) kit, commercial ELISAs using MESACUP Mitochondria M2 kit (old-M2 ELISA) and new-M2 ELISA, and immunoblotting on bovine heart mitochondria. RESULTS: Each test gave the following positive results; antimitochondrial antibodies (AMA) by immunofluorescence in 71 (87%) out of the 82 sera, enzymatic inhibitory antibody to pyruvate dehydrogenase complex (PDC) by EMA in 61 (74%), immunoglobulin (Ig) G class anti-PDC antibody by old-M2 ELISA in 55 (67%), IgG/M/A class anti-E2 subunit of PDC (PDC-E2)/anti-E2 subunit of branched chain oxo-acid dehydrogenase complex (BCOADC-E2)/anti-E2 subunit of 2-oxoglutarate dehydrogenase complex (OGDC-E2) antibodies by new-M2 ELISA in 73 (89%), and IgG, IgM, or IgA class antibodies against at least one of the 2-OADC enzymes by immunoblotting in 82 (100%). Fifty-three of the 82 sera (65%) were all positive by these five assays. Of the 18 sera that were positive by new-M2 ELISA but negative by old-M2 ELISA, 12 were theoretically interpretable. Of the 11 sera that were negative for AMA by immunofluorescence but positive for at least one of anti-2-OADC enzymes by immunoblotting, four (36%) were positive by new-M2 ELISA, whereas only two and one sera were positive by EMA and old-M2 ELISA, respectively. CONCLUSIONS: Our results indicated that the sensitivity of the newly developed new-M2 ELISA was higher than that of EMA and old-M2 ELISA, and comparable with that of immunofluorescence. However, it is still unclear whether the new-M2 ELISA could replace the conventional immunofluorescence testing for routine assay requests because six (7%) sera showed discrepant results between these two assays.     

IFA和ELISA在肿瘤患者血清SARS-CoV抗体测定中的对比分析

目的探讨酶联免疫吸附试验(ELISA)和间接免疫荧光法(IFA)检测严重急性呼吸综合征(SARS)冠状病毒(SARS-CoY)&体在SARS病原中诊断中的特异性及其在肿瘤患者血清中的假阳性问题。方法 应用ELISA和IFA检测了111例正常对照和40例肿瘤患者血清中SARS-CoV抗体的阳性率。结果在111例正常对照和40例肿瘤患者中,IgM抗体均阴性,IgG抗体在正常对照中的阳性率为3.6%(4/111),在肿瘤患者中的阳性率17.5％(7/40),IgC抗体诊断SARS的特异性为96.4％,两种抗体同时阳性诊断SARS的特异性为100％。经IFA检测,上述SARS-CoV抗体阳性者均为阴性。结论 IFA诊断SARS的特异性为100％,ELISA诊断SARS存在一定的假阳性。应用ELISA测定SARS-CoV抗体时,应同时测定两种抗体以降低诊断的假阳性率,提高诊断的特异性。     

Evaluation of quaternary aminoethyl-Sephadex A50 column chromatography for detection of anti-cytomegalovirus immunoglobulin M

The sensitivity of an indirect fluorescent antibody (IFA) test (screening test) for the detection of antibodies to cytomegalovirus (CMV) was examined by using 128 serum specimens and quaternary aminoethyl (QAE)-Sephadex A50 column chromatography to separate IgM from IgG class antibodies. Of these 128 rheumatoid factor-negative serum samples, only 54 (42%) continued to produce fluorescence after the IgG was removed by the Sephadex column (yielding a theoretical 1:10 dilution) and the samples were retested for IgM (confirming test). Of the 54 specimens that were positive on both the screening and confirming IgM IFA assays, 53 (98%) had titers of 1:80 or more. Of the 74 sera that were negative on the confirming test, only 10 (14%) had titers of 1:80 on the initial screening test, and none was greater than 1:80. Immunoglobulin measurements of eight serum samples before and after column treatment indicated excellent IgG removal but poor IgM recovery (a mean loss of 85% of the total IgM). Comparison of the time needed to detect IgM in CMV culture-positive patients demonstrated an increase from 17.1 to 28.3 days (serum tested by IFA before and after column separation, respectively). Therefore, confirmation of anti-CMV IgM in serum specimens by QAE-Sephadex A50 column chromatography seems to be of low sensitivity.     

Utility of an immunocapture-agglutination test and an enzyme-linked immunosorbent assay test against cytosolic proteins from Brucella melitensis B115 in the diagnosis and follow-up of human acute brucellosis

The utility of an immunocapture-agglutination (Brucellacapt, Vircell SL, Granada, Spain) test and an enzyme-linked immunosorbent assay IgG, IgA, and IgM (ELISA-IgG, ELISA-IgA, ELISA-IgM) against cytosolic proteins from Brucella melitensis B115 (R) was compared with ELISA-IgG, ELISA-IgA, and ELISA-IgM against smooth lipopolysaccharide (S-LPS) from B. melitensis 16M (S), serum agglutination test (SAT), and Coombs test in the diagnosis and follow-up for 10 months of 51 patients with acute brucellosis. The sensitivities of ELISA tests against cytosolic proteins varied from 49.0 % for ELISA-IgG to 64.7% for ELISA-IgM and were lower than the sensitivities showed by ELISA S-LPS (from 88.2% to 92.2%), SAT (88.2%), Coombs (96.1%), and Brucellacapt (98.0%) tests. Specificity was over 93% in all cases. The evolutionary behavior of the SAT, Coombs, and Brucellacapt tests was similar. There was a decrease of between 20% and 40% in antibody titer in the 10th month of evolution after treatment. The evolutional curves of IgG, IgA, and IgM against cytosolic protein increased slightly till the eighth month. The specific IgM and IgA antibodies against protein fractions began to show a drop from the eighth month on, showing levels slightly lower than the initial sera values by the end of the 10th month. In this month, titers of specific IgG against proteins fractions remained higher than the titers showed by the initial sera.