Seroepidemiology of enteric fever and brucellosis in a developing country

The seroepidemiology of infection due to S. typhi and Brucella spp. in a sub-tropical, developing country of the Middle East was investigated in 130 children of age less than 1 to 15 years, 117 young adults (16-30 years) and in a further 40 adults (greater than 30 years) using an agglutination test to detect antibodies to of S. typhi, B. abortus and B. melitensis. Only 4.5% of the children's and adult's sera respectively showed the presence of antibodies to both O and H antigens of S. typhi with reciprocal titers greater than or equal to 80. Prevalence rates of 11.6%, 11.3% respectively in children and adults of antibodies to B. abortus with reciprocal titers greater than or equal to 80 and similarly 13.1%, 12.4% to B. melitensis were found. The usefulness of a single Widal test to diagnose enteric fever in a non-endemic area and the agglutination test in the diagnosis of brucellosis in an endemic area was investigated. Of 244 patients with suspected enteric fever, the sera of 100 showed reciprocal antibody titers 80 to both O and H antigens and included in these were 8 patients with concurrent culture positivity. Among 138 children and adults with suspected brucellosis, 29 culture proven cases where serologically confirmed and a further 90 were detected serologically. The sera of all the culture proven cases exhibited antibody reciprocal titer greater than or equal to 640. A similar antibody response was noted in seventy-two of the culture negative cases diagnosed serologically for brucellosis. In culture proven, cases B. melitensis was the isolate. Comparative evaluation indicated the potential usefulness of the Rose Bengal Antigen Slide Agglutination Test (Brucelloside Test) as a screening test in the serodiagnosis of human brucellosis.     

Abnormal anti-Epstein Barr virus antibodies in carriers of the X-linked lymphoproliferative syndrome and in females at risk

The asymptomatic hemizygous female carriers of the X-linked lymphoproliferative syndrome (XLP) have abnormal antibody responses to EBV. This suggests partial expression of the defect that leads to EBV-provoked life-threatening diseases in their affected sons. EBV specific antibodies were measured in 65 serum samples of 12 obligate carrier females and seven of their daughters (females at risk) during periods ranging from 1 to 5 yr. Abnormal qualitative antiviral capsid antigen (VCA) IgG titers were nearly fourfold higher than normal controls, two carriers had persistent IgM anti-VCA antibody, two-thirds had persistent IgA anti-VCA antibody, and half of the women had titers to early antigen (EA). Five of seven females exhibited a similar persistent pattern. In contrast, none of the unaffected family members nor 23 normal controls expressed IgA or IgM titers to VCA even with high exposure to the virus, and anti-EA was detected in only one control. Therefore, these findings may prove useful for detecting carriers of the syndrome. Abnormal anti-EBV titers similar to the carrier pattern have been reported in patients and other immunosuppressed individuals, and are indicative of active viral infection.     

用PO4细胞为靶细胞检测人血清中Epstein-Barr病毒IgA/MA抗体以诊断鼻咽癌

近年来,常用未经丙酮固定的、由丁酸和巴豆油激活的B95-8细胞或P3HR-1细胞为靶细胞,检测人血清中EB病毒IgA/MA抗体以早期诊断鼻咽癌,效果良好。但由于B95-8细胞含有多种EB病毒抗原,不能用丙酮固定,需多次离心沉淀,在浮悬状态下检测,技术比较复杂。     

Comparison of heterologous adult Brugia malayi and homologous Onchocerca volvulus antigen in the enzyme-linked immunosorbent assay (ELISA) for Guatemalan onchocerciasis

To determine the susceptibility of a heterologous filarial antigen for measuring Onchocerca volvulus antibodies, worms were compared using an enzyme-linked immunosorbent assay (ELISA) test. Control serum samples from helminth-free U.S. residents and from helminth-infected but filariae-free Salvadoran residents were tested and compared with serum obtained from microfilariae-positive and -negative Guatemalan residents living in an area of endemic onchocerciasis. The results showed that none of the sera from U.S. residents had positive O. volvulus ELISA titers (greater than or equal to 1:160); however, 8.51% (4/47) had positive B. malayi ELISA titers (greater than or equal to 1:640). The geometric mean titers with the B. malayi ELISA test were higher than with the O. volvulus ELISA test--in sera from 47 U.S. residents (1:219 vs. 1:49), from 108 Salvadoran residents (1:92 vs. 1:71), and from 145 microfilariae-negative (1:539 vs. 1:167) and 303 microfilariae-positive (1:1,270 vs. 1:561) Guatemalan residents. The B. malayi ELISA test exhibited slightly less sensitivity than the homologous O. volvulus ELISA test; nevertheless, a good correlation (r = 0.74) was found between the 2 test antigens, indicating that the B. malayi antigen could be used to measure O. volvulus antibodies.     

IgA and IgM anti-Naegleria fowleri antibodies in human serum and saliva

Antibodies from IgA and IgM classes that recognize Naegleria fowleri (Nf) proteins were detected by the ELISA assay in serum and saliva from three groups of people: (i) subjects with upper respiratory tract infections (URTI) living in the parasite-endemic area, (ii) healthy persons from the same area, and (iii) healthy persons from a parasite-nonendemic area. In serum and in saliva the titers of IgA antibodies to Naegleria fowleri in the group of patients with URTI was significantly higher than that of the healthy group in the parasite-endemic area; also the titers of IgM antibodies in serum were significantly higher in patients. On the contrary, in saliva the antibodies were higher in healthy people from the parasite-endemic area. In all cases the subjects from the parasite-nonendemic area had lower antibody titers in serum and saliva.     

Antibody titers by mixed agglutination to varicella-zoster, herpes simplex and vaccinia viruses in patients with multiple sclerosis.

Antibody titers to varicella-zoster (V-Z), herpes simplex (HSV) and vaccinia viruses were determined in sera collected in Buffalo, NY, from patients with multiple sclerosis (MS), patients with other diseases, and normal individuals. The mixed agglutination test employing cell cultures infected with one of the above viruses as a source of antigen was used to determine antibody titers. The geometric mean titer (GMT) for V-Z in 59 MS cases was significantly higher than that observed in patients with other diseases and in normal individuals. The GMT for vaccinia was also higher in MS patients but the difference was not as great as for V-Z. No difference was observed in the titer of antibodies for HSV. Similar results were obtained for 51 MS patients compared to healthy controls matched for sex and age. Higher antibody titers for V-Z and HSV were observed in cerebrospinal fluids from MS cases than in those from non-MS CNS patients in Baltimore, MD. Antibodies to V-Z, HSV and vaccinia were detected in washing fluids from brain homogenates of MS cases.     

Comparison of antibodies to Leptospira in white-tailed deer (Odocoileus virginianus) and cattle in Ohio

A survey was conducted to determine the prevalence of leptospiral antibodies in sera from 248 white-tailed deer (Odocoileus virginianus) in Ohio. The sera were collected at check stations during the hunting season in 1983. The microscopic agglutination microtiter test was used to determine the presence of antibodies to Leptospira interrogans serovars pomona, icterohemorrhagiae, canicola, hardjo, and grippotyphosa. Eighteen of 248 (7.3%) serum samples had antibody titers (greater than or equal to 1:100) to at least one of the five serovars tested, with three of these samples reacting to more than one serovar. Prevalence did not differ significantly between sex or age groups. The serovar antigens reacting most frequently with serum antibodies were grippotyphosa (10 of 22, 45.5%) and pomona (eight of 22, 36.4%). Sera agglutinating with pomona antigen had higher titers (ranging from 1:200 to 1:6,400) than did sera agglutinating with the other serovars. These results were compared to results obtained from cattle tested at the Ohio Department of Agriculture Laboratories during 1983. There was a significant relationship between pomona infections detected in deer and cattle (P less than 0.05), but not with grippotyphosa.     

血清中Epstein-Barr病毒膜抗原IgA抗体检测法的改进及应用

用萄葡球菌菌体A蛋白(SPA)预先处理被检血清,以去除抗体IgG部份的竞争性结合,提高了间接免疫荧光法检查鼻咽癌病人血清中EB病毒膜抗原IgA(IgA/MA)抗体的敏感性及特异性,检查48例鼻咽癌病人血清IgA/MA抗体,阳性率为100％,血清几何平均滴度为141;40例其它恶性肿瘤病人和46例正常人都检不出IgA/MA抗体,免疫荧光法测得IgA/MA抗体阴性的6例鼻咽癌病人血清,SPA吸收后呈阳性反应,此改进方法可用以追踪观察鼻咽癌病的病程及预后。     

Antibodies against Toxoplasma gondii in the Pacific bottlenose dolphin (Tursiops aduncus) from the Solomon Islands

Serum samples from 58 Pacific bottlenose dolphins (Tursiops aduncus) from the Solomon Islands were tested for the IgG antibody to Toxoplasma gondii by the latex agglutination test (LAT), enzyme-linked immunosorbent assay (ELISA), and immunoblotting. The ELISA cut-off value was taken as OD > or = 0.276, and the final dilution ratio, recognized as positive, was represented by the end titer. In 25 of 58 samples, no antibody activity was detected by LAT and ELISA. In 8 of 58 samples, anti-T. gondii IgG antibodies were detected by both LAT and ELISA, with titers of greater than 1 : 64 and 1 : 160, respectively. By immunoblotting, the 8 serum samples producing higher titers showed specific antibody IgG binding to several antigens on the T. gondii lane, but not on the Neospora caninum lane. No specific bands were noted on the lanes for either parasite in the 25 serum samples for which no antibody activity was detected. The specific binding of IgG antibodies to T. gondii antigens observed for serum samples producing higher titers suggests that Pacific bottlenose dolphins from the Solomon islands are exposed to T. gondii.     

Increased concentrations of antibody against heat shock protein in patients with myeloperoxidase anti-neutrophil cytoplasmic autoantibody positive microscopic polyangiitis

To determine serum antibody against human and bacterial heat shock protein (HSP) 60/70 in myeloperoxidase (MPO)-specific anti-neutrophil cytoplasmic autoantibody (ANCA) positive microscopic polyangiitis (MPA), 58 patients with MPO-ANCA positive MPA, 48 with RA (rheumatoid arthritis) and 40 with SLE (systemic lupus erythematosus) were studied. Serum antibodies against HSP (human HSP 70, human HSP 60, Mycobacterium HSP 70, and Escherichia coli HSP 60) were measured by sandwich ELISA. The frequency of anti-human HSP 60/70 antibody positive patients was significantly greater in MPO-ANCA positive MPA than SLE and healthy controls. Anti-human HSP 60/70 antibody titers in patients with MPO-ANCA positive MPA were significantly higher than those of healthy controls; anti-bacterial HSP 60/70 antibody titers were also higher. There was a significant correlation between titers of anti-human HSP 70 antibody and anti-Mycobacterium HSP 70 antibody. A correlation was also found between titers of anti-human HSP 70 antibody and anti-human HSP 60 antibody. Anti-human and bacterial HSP 60/70 antibody titers changed in parallel with disease activity in patients with antibody positive MPA. The anti-HSP antibody titer was also increased in patients with RA and SLE. These results suggest that an immunological background via anti-HSP 60/70 antibodies might be associated with pathogenesis in MPO-ANCA positive MPA.     

Antibody-dependent cellular cytotoxicity-inducing antibodies against human immunodeficiency virus. Presence at different clinical stages

The presence of antibodies mediating antibody-dependent cellular cytotoxicity (ADCC) against human immunodeficiency virus (HIV)-infected target cells was investigated with 170 sera from patients with varying severity of HIV infection. Approximately 40% of sera from individuals representing all stages of infection were ADCC-positive when tested against HTLV-IIIB infected 0937 clone 2 target cells. The positive sera had higher HIV antibody titers as measured by enzyme-linked immunosorbent assay compared with ADCC-negative sera. ADCC titers were lower in patients with acquired immune deficiency syndrome than in asymptomatic carriers. This decline in ADCC titer was not correlated with a general decrease of HIV antibodies. No correlation between the CD4:CD8 lymphocyte ratio and ADCC activity was found. The possible beneficial effect of ADCC-inducing antibodies early in infection is discussed in relation to the effect of ADCC-inducing antibodies in other retrovirus systems and to the nature of lentivirus infections.     

Inactivated SARS-CoV vaccine prepared from whole virus induces a high level of neutralizing antibodies in BALB/c mice

We tested the ability of inactivated SARS-CoV vaccine to induce neutralizing antibodies in BALB/c mice. The inactivated vaccine was prepared by SARS-CoV virus propagation in Vero cells, with subsequent beta-propiolactone inactivation and Sepharose 4FF column chromatography purification. One hundred forty BALB/c female mice were divided into seven groups of 20 mice each. Of the seven groups, three groups were inoculated with 0.1, 1, and 3 microg of the vaccine without adjuvant while three other groups were inoculated at the same three dosages of vaccine with aluminum hydroxide as adjuvant, respectively. The remaining group was set up as a blank control. Each mouse was inoculated twice at an interval of 3 weeks. One week after the second immunization, mice sera were collected to detect serum neutralizing antibodies. An assay for determining neutralizing antibody titers was developed. The results can be summarized as follows: (1) higher dosages of vaccine induced higher levels of neutralizing antibody titer; (2) the level of neutralizing antibodies induced by the inoculation with aluminum hydroxide adjuvant was slightly higher than that without adjuvant, but the difference was not statistically significant.     

Development and Evaluation of a Simple Latex Agglutination Test for Diagnosis of Tuberculosis

A simple latex agglutination test (SLAT) based on modifications of existing serodiagnostic techniques, in which commercially available reagents are used, was developed for detection of antibodies against Mycobacterium tuberculosis. Tests performed on 553 serum samples from 316 individuals, including 117 bacteriologically confirmed active tuberculosis patients, showed 80% positive titers. Sera from 12 patients with arrested tuberculosis showed 91% positive titers. Nonspecific reactions were noted in 5% of 160 serums from selected normal individuals and patients with diseases other than tuberculosis. The antibodies detected by the SLAT method were found to be relatively stable when exposed to low temperatures, whereas high temperatures reduced the antibody titer considerably. Disodium ethylenediaminetetraacetic acid inactivation of serum complement was found to be satisfactory. No variation of tuberculosis antibody titer was noted in tests on multiple specimens from patients whose conditions were stabilized. However, considerable fluctuation was encountered in antibody titers obtained on recently detected individuals. Data obtained in this study indicate that the modified procedures of the SLAT method could replace the tuberculin skin test for simple screening of tuberculosis in adults.     

Persistence of antibodies of the IgG class against cytomegaloviruses in blood donors

High titre plasmas gained from blood donors are the initial material used for producing human immunoglobulin containing cytomegalovirus antibodies (CMV-HIG). For this purpose the sera gained from 467 permanent donors at the District Institute for Blood and Transfusion Service in Berlin were investigated on their CMV antibody content of IgG class by means of an indirect immunofluorescence test (IFT). The infection rate of blood donors amounted to 62% (291/467). CMV-IgG titre greater than or equal to 1:40 was determined in 88 sera (18.8%) and greater than or equal to 1:160 in 14 sera (3%). Two CMV-HIG laboratory samples (charges 3113 and 3117) were produced from these plasmas. Particularly immunoglobulin fraction of charge 3117 (No. 3117 N II) revealed excellent antibody titres (IFT 1:640 [CMV-IgG] or 1:40 [CMV-IgM] respectively, neutralisation test 1:32). Checks made with the donors' CMV-IgG titres after repeated application of plasmapheresis resulted in maximal titres changes of two dilution stages in a period of 15 months. Thus, in producing CMV-HIG a sure, tested pool of donors can be resorted to.     

Epidemiological studies on echovirus type 18 infection in Toyama Prefecture

Echovirus type 18 (echo 18) was isolated from six aseptic meningitis children in Fukumitsu-machi, Toyama Prefecture, from July to August, 1980. This was the first virologically-confirmed epidemic of aseptic meningitis due to echo 18 in Japan. Epidemiological studies on the prevalence of this virus among the inhabitants in Toyama Prefecture were also performed. The results are summarized as follows. (1) Significant increases in neutralizing antibody titers against echo 18 were observed in all the paired sera of aseptic meningitis patients from whom echo 18 was isolated. (2) In October, 1980, echo 18 was also isolated from healthy children or from infants suffering from gastroenteritis in other areas of Toyama Prefecture. (3) Among the sera collected from 50 children (aged smaller than or equal to 12) in Fukumitsumachi in December, 1980, neutralizing antibodies against echo 18 were detected only in the younger groups (aged smaller than or equal to 8), 58.9% of these age groups showing a titer of 4 or higher. (4) Another epidemic around 1963 by echo 18 in Toyama Prefecture was retrospectively suggested by the examinations of sera collected in 1978 and 1981 from inhabitants in various areas of Toyama Prefecture. (5) Neutralizing antibody titers against the strain isolated, No. 35'80, were significantly higher than those against strain 'Metcalf,' the prototype of echo 18, in most sera including both aseptic meningitis patients and healthy inhabitants.     

Studies on the presence of antibodies to EB virus and other herpesviruses in normal children and in infectious mononucleosis.

Children free from infectious disease have been examined by indirect immunofluorescence for the presence of antibodies to the intracellular capsid antigen of Epstein-Barr virus (EBV). In the first year of life 46%, between 2 and 6 years of age 66%, and between 7 and 14 years 91%, of the children proved positive. The corresponding percentages for the presence of antibodies to cytomegalovirus (CMV) and herpes simplex virus were about 50%, irrespective of the children's age. Serum samples from 69 patients suffering from infectious mononucleosis (IM) were tested for anti-EBV antibodies. Of the 29 Paul-Bunnell-positive patients 22 had antibodies, 11 of them in high titres (greater than 1 : 80). Of the 40 Paul-Bunnell-negative cases only 21 had antibodies, 8 in high titres. Of the Paul-Bunnell-negative cases, 73% were found to have anti-CMV antibodies, 32% in high titre. The respective percentages for the Paul-Bunnell-positive cases were 42% and 10%.     

儿童传染性单核细胞增多症嗜异性凝集抗体与EBV-VCA-IgM检测的比较

目的比较分析外周血异型淋巴细胞（AL）、血清嗜异性抗体（HA）及EBV-VCA-IgM在儿童传染性单核细胞增多症（IM）诊断中的意义。方法对238例儿童IM进行AL计数、AVITEX-IM乳胶凝集法检测HA、ELISA法检测EBV-VCA-IgM,分析比较此3种检验方法的阳性率。结果（1）82．35％的患儿外周血AL〉10％,其中71．94％在病程第2周AL〉10％。（2）HA阳性者占48．85％,HA阳性率〈3岁（14．55％）明显低于≥3岁（53．90％）,P〈0．05。（3）EBV-VCA-IgM 163例阳性（75．81％）,各年龄组EBV-VCA-IgM阳性率差异无显著性。（4）72例同时检测血清EBV-VCA-IgM和HA,EBV-VCA-IgM阳性者58例（80．56％）,HA阳性者36例（50％）,EBV-VCA-IgM与HA均为阳性者33例（45．83％）,EBV-VCA-IgM的阳性检出率明显高于HA。结论EBV-VCA-IgM的阳件率明昂高干HA的阳件塞罩期殛时拱奉台EBV抗体对于及早、快速、准确诊断IM十分必要。     

鼻咽癌患者血清中抗Epstein-Barr病毒早期和晚期膜抗原抗体的检测

对Epstein-Barr(EB)病毒抗原的研究,发现有淋巴细胞确定的膜抗原(Lydma)、早期抗原(EA)、壳抗原(VCA)、核抗原(EBNA)、早期膜抗原(EMA)和晚期膜抗原(LMA)。除了Lydma抗原外,鼻咽癌患者对上述抗原均产生相应的IgG和IgA抗体。因而研究这些抗体,对阐明EB病毒与鼻咽癌的关系及鼻咽癌的早期诊断都十分有价值。     

A preclinical study comparing approaches for augmenting the immunogenicity of a heptavalent KLH-conjugate vaccine against epithelial cancers

Previously using a series of monovalent vaccines, we demonstrated that the optimal method for inducing an antibody response against cancer cell-surface antigens is covalent conjugation of the antigens to keyhole limpet hemocyanin (KLH) and the use of a saponin adjuvant. We have prepared a heptavalent-KLH conjugate vaccine containing the seven epithelial cancer antigens GM2, Globo H, Lewis(y), TF(c), Tn(c), STn(c), and glycosylated MUC1. In preparation for testing this vaccine in the clinic, we tested the impact on antibody induction of administering the individual conjugates plus adjuvant compared with a mixture of the seven conjugates plus adjuvant, and of several variables thought to augment immunogenicity. These include approaches for decreasing suppressor cell activity or increasing helper T-lymphocyte activity (low dose cyclophosphamide or anti-CTLA-4 MAb), different saponin adjuvants at various doses (QS-21 and GPI-0100), and different methods of formulation (lyophilization and use of polysorbate 80). We find that: (1). Immunization with the heptavalent-KLH conjugate plus GPI-0100 vaccine induces antibodies against the seven antigens of comparable titer to those induced by the individual-KLH conjugate vaccines, high titers of antibodies against Tn (median ELISA titer IgM/IgG 320/10240), STn (640/5120), TF (320/10240), MUC1 (80/20480), and globo H (640/40); while lower titers of antibodies against Lewis(y)()(160/0) and only occasional antibodies against GM2 are induced. (2). These antibodies reacted with the purified synthetic antigens by ELISA, and with naturally expressed antigens on the cancer cell surface by FACS. (3). None of the approaches for further altering the suppressor cell/helper T-cell balance nor changes to the standard formulation by lyophilization or use of polysorbate 80 had any impact on antibody titers. (4). An optimal dose of saponin adjuvant, QS-21 (50 microg) or GPI-0100 (1000 microg), is required for optimal antibody titers. This heptavalent vaccine is sufficiently optimized for testing in the clinic.     

Use of the indirect hemagglutination reaction for detecting antibodies to Pseudomonas pyocyanea in acute suppurative destructive pneumonia]

No less than 4-fold increase in the antibody titres to Pseudomonas aeruginosa during the infectious process served as laboratory confirmation of its participation in the infectious process. In 49 of 91 patients hemagglutining titre exceeded the diagnostic one (1:640). In 9 patients (chiefly in infants) hemagglutinin titre remained low, but there was a rise of antibody level to Pseudomonas aeruginosa during the disease. High hemagglutinin titres were noted in 12 patients on admission to the clinic, with reduction of the antibody titres at the late periods of the disease. Antibody titres remained unchanged during the disease in 15 patients. In 3 cases the indirect hemagglutination test was assesed as negative. In the rest of the patients hemagglutinin titres varied within the range of the diagnostic titre. Thus, the indirect hemagglutination test with erythrocytic diagnostic agent permitting to determine antibodies to Pseudomonas aeruginosa could be used at the clinic in combination with bacteriological and other investigations for establishing etiology of the destructive process.