Comparative study of cell-immunoenzymatic methods for the estimation of IgG and IgM anti-brucella antibodies in the diagnosis of human brucellosis

Two methods which employ whole cells are described and compared for the detection of human IgG and IgM anti-brucella antibodies. Dot ELISA and ELISA were shown to be suitable for a screening diagnosis of human brucellosis. Titres of antibodies obtained by dot ELISA showed 100% coincidence for IgG and 97% for IgM, compared with agglutination and complement fixation tests; when ELISA was used 11 % positive sera were not detected. The comparison of these two methods with the conventional serological test kit indicated that both dot ELISA and ELISA were sensitive, reproducible and specific for the quantification of IgG and IgM anti-brucella antibodies.     

Comparative study of cell-immunoenzymatic methods for the estimation of IgG and IgM anti-Brucella antibodies in the diagnosis of human brucellosis

Two methods which employ whole cells are described and compared for the detection of human IgG and IgM anti-brucella antibodies. Dot ELISA and ELISA were shown to be suitable for a screening diagnosis of human brucellosis. Titres of antibodies obtained by dot ELISA showed 100% coincidence for IgG and 97% for IgM, compared with agglutination and complement fixation tests; when ELISA was used 11% positive sera were not detected. The comparison of these two methods with the conventional serological test kit indicated that both dot ELISA and ELISA were sensitive, reproducible and specific for the quantification of IgG and IgM antibrucella antibodies.     

替加环素与米诺环素治疗布鲁菌病的进展

布鲁菌病是由布鲁菌引起的人畜共患传染性疾病，由带菌动物通过多种途径传染给易感人群，可侵犯人体各器官系统。目前推荐的抗布鲁菌治疗方案为多西环素、利福平、喹诺酮类、氨基糖类等抗菌药物组合的二联或者三联抗菌方案，但治疗无效率和疾病复发率仍较高。替加环素和米诺环素作为四环素类药物，经体外实验证明均可有效抑制布鲁菌，且50%最小抑菌浓度（50% minimal inhibitory concentration，MIC50） 和 90%最小抑菌浓度（90% minimal inhibitory concentration，MIC90）均较低。临床研究亦证实，包含替加环素和米诺环素的联合抗菌方案具有较低的治疗无效率和疾病复发率。因此，替加环素和米诺环素可作为治疗布鲁菌病的候选用药。本文就布鲁菌病传统治疗方案以及替加环素和米诺环素的抗菌效果进行综述。     

自身免疫性肝炎合并慢性布鲁菌病1例并文献复习

目的探讨自身免疫性肝炎合并布鲁菌感染时的临床特征、诊断及其治疗等特点。方法回顾性分析我院确诊的1例自身免疫性肝炎合并慢性布鲁菌感染患者的临床特点、相关指标的检测结果并结合文献报道进行总结。结果患者临床表现缺乏特异性。血培养和血清抗体检测呈阳性,C-反应蛋白(CRP)可见升高而降钙素原(PCT)变化不明显。结论布鲁菌病不能仅从临床表现来鉴别和诊断,更不能因为缺少流行病学资料而否定。自身免疫性肝炎患者的CRP水平对急性炎症的敏感性会降低,但仍可以作为疗效判断的参考指标。应适度增加疗程,定期复查,以更好的控制慢性布鲁菌感染。     

Australia-SH Antigen in Hepatitis Patients in London

Australia-SH antigen was found in 11 out of 27 sera (40%) obtained from patients with acute viral hepatitis soon after their admission to hospital. Six out of 11 positive sera were collected within the first 12 days of illness; die remaining five were collected between the 13th and 30th days. The antigen was detected by immunodiffusion in agarose gel, five different indicator sera containing Australia-SH antibodies being used. The specificity of these sera was found to be very similar.All of the patients'' sera were examined independently by two separate laboratories. The results obtained by the laboratories were in close agreement, though the techniques varied in detail.     

Depression of Cellular Immunity in Pregnancy due to a Serum Factor

Lymphocytes from pregnant women and non-pregnant individuals were cultured under the stimulus of phytohaemagglutinin in the presence of their own and heterologous (allogeneic) sera. The results indicate that heterologous sera have an inhibitory effect on the lymphocyte transformation rate and suggest that the inhibitory property is more powerful in pregnant and fetal sera. Conversely, the addition of heterologous non-pregnant sera to cultures of pregnant lymphocytes increases their transformation rate. These findings suggest that there is a serum inhibitor in pregnancy and this finding may be relevant to the survival of the fetal allograft.     

Type-specific pneumococcal agglutinating sera in precipitation tests

Diagnostic agglutinating sera to pneumococci of different serotypes have been studied with respect to their capacity for taking part in the reactions of precipitation with capsular pneumococcal polysaccharides. The sera have proved to be highly active and specific in the reactions of double immunodiffusion and immunoelectrophoresis. The sera under study have also been found to react with cattle serum, one of the components of the medium used for the cultivation of pneumococci.     

Preservation of Coombs' serum in liquid nitrogen

Experiments with Coombs-sera as raw and usable sera having different levels of titre were carried out in order to examine their stability in fluid nitrogen. Therewith, significant differences concerning the storing stability did not appear. Generally seen two third of the sera retained their initial titres at the end of one year in store.     

Problems in the preparation and testing of anti-human globulin sera with regard to their use

The problems in manufacturing and testing A.H.G. sera with regard to their application are presented. The sera offered at present do not answer the requirements wanted. Therefore, the properties of the A.H.G. sera to be demanded in practice are presented and the methods of manufacture, the criteria of testing them, and the results thereof, as well, are presented.     

Effects of steroid hormones in fetal bovine serum on plating ang cloning of human cells in vitro.

Fetal bovine sera from each of three different commercial sources were tested for their ability to support cloning of human fibroblastoid cells in vitro. Cloning efficiencies varied according to serum source. Serum (10 samples) from company A did not support growth, while sera (10 samples) from companies B and C provided adequate to excellent conditions for cloning and growth. Cells from neonatal foreskin or embryonic lung responded to each serum similarly. Bovine serum albumin type H7 from company C supported cell growth in media without serum. Sera containing 1.0 ng per ml or more of progesterone inhibited growth, whereas sera containing less than 1.0 ng per ml supported cloning and growth. In the low progesterone sera, the concentration of 17-beta-estradiol exceeded 100 pg per ml. Growth supporting sera could be made non-supportive by adding 0.1 mug per ml of progesterone. The addition to non-supporative sera of 0.1 mug per ml of 17-beta-estradiol or hydrocortisone made these sera supportive of cell growth. Addition of estrogen or hydrocortisone to a culture medium that inhibits growth, with subsequent reversal of the inhibitory effect, implies that these hormones competitively regulate growth of responsive cells in vitro.     

Bone Marrow Colony-stimulating Activity of Sera in Infectious Mononucleosis

From 44 to 100% of sera from patients with infectious mononucleosis exhibited the capacity to stimulate colony formation in vitro by mouse bone marrow cells. The proportion of sera with colony-stimulating activity was highest in patients with a short fever period and developing low Paul-Bunnell titres. Patients with a more severe course of the disease generally displayed no, or only weak, colony-stimulating activity in their sera, and also had higher Paul-Bunnell titres. The level of serum colony-stimulating activity tended to fall in the convalescent stages of the disease.     

Identification of centromere proteins in different mammalian cells

The characterization of centromeric proteins is facilitated using anti-centromere antibodies present in the sera of patients with the CREST variant of scleroderma. We have employed these sera to determine whether or not those proteins are present in different mammalian species, as well as to study their tissue distribution. Here, we describe the immunofluorescent pattern and the proteins recognized by CREST sera in dividing and resting cells from mouse, rat, swine, hamster, rabbit, and man. In nuclear preparations from cultured cells, thymocytes and spermatozoa from these species, the antigens recognized by CREST sera are proteins of 18 to 20 kDa in all species tested, except in rat. Additionally, two peptides of 80 and 140 kDa were observed in human preparations. In contrast, a 50 kDa peptide is the primary protein detected by the sera in rat nuclei.     

Immunochemical characterization of the anti-RNA antibodies found in scleroderma and systemic lupus erythematosus. I. Differences in reactivity with Poly (U) and Poly-(A) Poly (U).

In a previous study, all 40 sera from patients with scleroderma, 20 of 40 sera from SLE patients, but none of 40 sera from normal controls, were found to have antibodies to ssRNA. All scleroderma sera were also found to react with HSA-coupled uridine and UMP and their reaction with HSA-coupled uridine and UMP and their reaction with ssRNA could be inhibited by uracil, uridine, and UMP. To characterize further these uracil-specific anti-RNA antibodies found in scleroderma and compare them with the anti-RNA antibodies found in SLE, we tested their reactivity with Poly (U) and with Poly (A)-Poly (U) and all but one failed to react with Poly (A)-Poly (U). This same serum was the only one in which the reaction with Poly (U) could not be inhibited with uracil. Reactivity of SLE sera was strikingly different from that found in scleroderma sera. Seventeen of 34 SLE sera studied reacted with ssRNA but only four of these reacted with Poly (U). Conversely, two SLE sera that reacted with Poly (U) did not react with ssRNA. Fifteen reacted with Poly (A)-Poly (U) and only two of these failed to react with ssRNA. Five SLE sera which were reactive with ssRNA did not precipitate with Poly (A)-Poly (U). All SLE sera which reacted with Poly (U) could be inhibited with uracil, although less effectively than in scleroderma. Reactivity with Poly (A)-Poly )U) was not inhibited with uracil nor with adenosine. These findings confirm that antibodies to RNA that are found in scleroderma are directed to uracil and thus specific to ssRNA, whereas RNA antibodies found in SLE sera are heterogeneous and directed to either the base, to the site of union of the base and sugar moiety to the ribose backbone, or to the helical structure of double stranded RNA. These differences and the respective antigenic specificities of these anti-RNA antibodies found in scleroderma and SLE may be theoretically important.     

Occurrence of Antirodies to Group Specific Chlamydia Antigen in Finnish Sheep,Cattle and Horse Sera

A serological survey on the occurrence of group-specific chlamydial antibodies in random sera of Finnish sheep, cattle and horses was performed. The whole material consisted of 1347 serum samples, including 432 ovine, 454 bovine and 461 equine sera. The sera were sent to the laboratory for various serological tests during 1968–1972. Of the ovine sera 9.5%, bovine 12.8 % and equine 7.1 % showed a titer ≥ 1:16 in the complement fixation test. No definite geographic differences could be found in the distribution of the herds which showed positive results. The ubiquity of chlamydial infections in domestic mammals and their role as a cause of clinical diseases is discussed.     

A simple and rapid method of determining the titer of herpes simplex virus antibodies in a study by immunoenzyme analysis of a single serum dilution

The result obtained in the study of the possibility of using the method for the determination of the titer of antibodies to herpes simplex virus by EIA techniques in a single dilution of the serum under test are presented. This method is based on the determination of the optical density of the serum titer (rcut) in different groups of sera with the use of the assay system, permitting the evaluation of the positive results obtained in the determination of their final dilution. The results obtained with the use of this method showed that error was 50% for high-titer sera, 60% for medium-titer sera and 30% for low-titer sera.     

Platelet-activating factor and serum components from oestrous mice co-operate to mimic the activity of 'early pregnancy factor' in the rosette inhibition assay

When male mouse spleen cells were incubated with a combination of platelet activating factor (PAF, 1-0-alkyl-2-acetyl-sn-glycero-3-phosphocholine) and sera from female mice in oestrus, the cells displayed a markedly increased rosette inhibition titre (RIT) when subsequently tested in the rosette inhibition assay. Neither PAF nor oestrous mouse sera alone could induce this effect, the combined action was required. Lyso-PAF could not substitute for the PAF, nor could male mouse sera nor the sera from females in dioestrus or metoestrus substitute for the oestrous mouse serum requirement. Pro-oestrous mouse sera could replace oestrous mouse sera but were less effective in their dose-responses. Studies on the mechanism of action of the PAF and oestrous mouse serum components suggested that the PAF stimulated the production and release of soluble factors (termed S2 factors) which by themselves could induce increased RIT values when applied to fresh spleen cells. The PAF-stimulated cell populations were rendered refractory to the action of these S2 factors and did not display increased RIT values, unless oestrous mouse serum was added. This serum acted to reverse the refractory state, allowing the S2 factors to exert their effect, and so cells treated with PAF and oestrous mouse serum displayed increased RIT values.     

Anterior pituitary cell antibodies detected in Hashimoto's thyroiditis and Graves' disease

An immunofluorescence study using unfixed cryostat sections of rat pituitary glands was carried out on sera from 34 patients with Hashimoto's thyroiditis, 28 patients with Graves' disease, 10 patients with thyroid adenoma and 50 healthy subjects. After absorption of sera with rat liver tissues, 19 of 34 patients retained reactivity to anterior pituitary cell antibodies (PCA, 55.8%). On the other hand, immunofluorescence in anterior pituitary cells was faint and detected in only 2 of 28 patients with Graves' disease (7.1%) after absorption of their sera with rat liver aceton powder. A similar result was also obtained when PCA were compared in the sera of Hashimoto's thyroiditis and Graves' disease with high titers of thyroid microsomal autoantibodies. PCA were detected neither in the sera of patients with thyroid adenoma nor in the healthy subjects. The present study suggests that PCA were considerably more prevalent in Hashimoto's thyroiditis than in Graves' disease.     

Circulating immune complexes as possible cause for anticomplementary activity in humans with malignant melanoma

Summary Sera from 98 melanoma patients, 20 noncancer patients with immune complex-associated diseases, and 90 normal donors were analyzed for anticomplementary (AC) activity by the complement consumption method. Some of these sera were also tested for immune complex-like materials by the Raji cell radioimmune assay. In addition, serum samples from ten melanoma patients were analyzed serially to correlate the AC activity with clinical course. Significant levels of Ac activity were found in 45% of melanoma sera, 75% of nonmalignant immune complex-associated disease sera, and 10% of normal donors' sera. In some patients, AC activity decreased and became undetectable as their disease progressed. AC-negative serum samples taken from melanoma patients late in the course of disease when the tumor burden was large became anticomplementary when mixed with autologous or allogeneic serum samples taken earlier at the time of little or no tumor burden. The early serum samples contained antibodies against autologous tumor extracts, as shown by a complement fixation test. Absorption of early serum samples with cultured allogeneic melanoma cells reduced their ability to consume complement when mixed with autologous late serum samples, suggesting the presence of free antigen in the latter. The mixed samples of early and late sera and the sera positive in the complement consumption test contained heavy nonmonomeric IgG. Therefore, the AC activity of melanoma sera could be due to tumor-associated antigen-antibody complexes.     

Preventive properties of the blood sera from persons vaccinated with a Proteus vaccine made from soluble antigen complexes

In experiments of the passive protection of mice the protective properties of sera obtained from humans before and after their immunization with Proteus vaccine used as a monopreparation or in combination with staphylococcal toxoid and/or pyoimmunogen were studied. When introduced in a single subcutaneous injection, Proteus vaccine prepared from soluble antigenic complexes ensured an increase in the protective properties of sera. The second injection of the vaccine essentially enhanced the protective potency of the sera of the immunized donors. The therapeutic injection of Proteus vaccine ensured the essential increase of the protective properties of the sera. This increase could be experimentally detected within at least 25-30 days from the beginning of immunization. The immunization of volunteers with Proteus vaccine in combination with pyoimmunogen and adsorbed staphylococcal toxoid ensured the maximum increase of the protective properties of their sera.     

Inhibition of mitogen-induced lymphocyte proliferation correlated to anticomplementary activity in sera from melanoma patients

Summary Sera from 98 melanoma patients and 90 normal donors were analyzed for antibody (Ab) to melanoma extracts, melanoma-associated antigen (Ag) and anticomplementary (Ac) activities by the microcomplement consumption technique. Sera were also tested for their ability to inhibit mitogen(phytohemagglutinin: PHA)-ininduced blastogenesis of normal lymphocytes. The results of the complement consumption assays were correlated with the results of inhibition of PHA-induced blastogenesis. Of 98 melanoma sera, 22% were Ab-positive, 30% were Ag-positive, and 44% were Ac-positive, in contrast to only 6% Ab-positive, no Ag-positive, and 7% Ac-positive in 90 normal sera. Fifty-nine percent of melanoma sera were inhibitory to PHA-induced blastogenesis, as against 12% of normal donors' sera. Ac-positive melanoma sera were significantly more inhibitory than Acnegative melanoma sera. The inhibitory activity of Acpositive sera was potentiated by the simultaneous presence of detectable Ag activity and was diminished by detectable Ab activity. Presence of Ab or Ag activity alone did not correlate with the inhibitory activity of the melanoma sera. Increasing incidence of Ac activity and ability to inhibit PHA-induced blastogenesis was observed with increasing tumor burden or advancing clinical stage. Sera from patients who displayed a delayed cutaneous hypersensitivity reaction to 2,4-dinitrochlorobenzene (DNCB) were less inhibitory to PHA-induced blastogenesis than the sera from patients who did not respond to this contact allergen. Thus, Ac activity may be one of the circulating factors responsible for the immunosuppressive effect of cancer patients' sera.