Detection of circulating antibodies in patients affected by Chromoblastomycosis by Cladosporium carrionii using double immunodiffusion

The antibodies in sera of patients affected by Chromoblastomycosis are detected using the technique of double immunodiffusion and the mycelial somatic antigens and the culture filtrates antigens of Cladosporium carrionii. From the 13 sera tested 8 have given positive results. The fresh serum from a patient under treatment gives 2 bands, while fresh serum from a non-treated patient gives 3 bands. The titre of antibodies was also determined for the two fresh sera, having found 1/4 for the patient under treatment and 1/32 for the non-treated one.     

Serologic testing for symptomatic coccidioidomycosis in immunocompetent and immunosuppressed hosts

Serologic studies are an important diagnostic tool in the clinical evaluation and follow-up of persons with coccidioidomycosis. Numerous types of serologic tests are available, including immunodiffusion, enzyme immunoassay, and complement fixation. We conducted a retrospective review of the results of 1,797 serologic tests spanning 12 months from the onset of coccidioidomycosis in 298 immunocompetent and 62 immunosuppressed persons with symptomatic infection. Using the onset of symptoms as a reference point, we plotted the positive or negative serologic results over time for both groups. Compared with the immunocompetent group, immunosuppressed persons had lower rates of seropositivity for every type of test during the first year after onset of symptoms for coccidioidomycosis, although many results did not achieve statistical significance. Combining the results of these tests increased the sensitivity of the serologic evaluation in immunocompromised patients. Immunosuppressed persons have the ability to mount a serologic response to coccidioidomycosis, but in some circumstances, multiple methods may be required to improve detection.     

应用单向放射免疫扩散法测定流感疫苗血凝素含量的研究

分析了单向放射免疫扩散法快速、准确测定流感疫苗血凝素含量的可行性。利用在打孔器孔径一定的条件下抗原浓度只与扩散圈直径的平方成线性关系,而与打孔器无关,测定流感疫苗血凝素含量。结果可见,流感疫苗3个型别血凝素扩散圈直径的平方均与其浓度成线性,r均大于0.99,变异系数均小于1%,平均回收率99.7%。单向放射免疫扩散法方法简便、快速、准确,适用于流感疫苗血凝素含量的测定。     

Radioimmunological study of the surface protein of the human serum low-density lipoprotein: comparison of the native particle and the products obtained by tryptic treatment.

The present report describes radioimmunological studies of the native low-density lipoprotein from human serum, and of the products obtained by limited tryptic treatment, i.e.a protein-depleted particle lacking some 20% of the original protein moiety and a peptide fraction of low molecular weight (<5000). The liberated peptides were highly immunogenic and elicited antibodies which reacted with both the native and protein-deficient lipoprotein particles. Moreover these peptides exhibited competitive reactivity with [¹²⁵I]-labelled low-density lipoprotein in binding with homologous antisera, and with antisera to the native and trypsin-treated lipoproteins. These findings suggest that the peptides liberated from low-density lipoprotein by tryptic digestion contain the major antigenic site(s) of the molecule. Consideration of the nature of the competitive displacement of radiolabelled low-density lipoprotein from antisera to low-density lipoprotein, to the trypsinised lipoprotein and to the peptide fraction indicate that a marked repetition of the antigenic site(s) occurs in the structure of the protein moiety, a possibility consistent with the recurrence of similar subunits in the apoprotein of low-density lipoprotein.     

Synthesis and bioactivities of poly(ethylene glycol)–chitosan hybrids

Poly(ethylene glycol)–chitosan hybrids of various molecular weights having different degree of substitution were synthesized, by reductive N-alkylation of chitosan with poly(ethylene glycol) aldehyde, to study their bioactivities. The influence of these chitosan derivatives on the reactive oxygen species generation from canine polymorphonuclear leukocyte cells was investigated in vitro by chemiluminescence response. Reactive oxygen species generation by the influence of poly(ethylene glycol)–chitosan hybrids was decreased with the increase of degree of substitution. The reduction of interaction of poly(ethylene glycol)–chitosan hybrids with polymorphonuclear leukocyte cells might be caused by the decrease of amino group in chitosan main chain and increase of the steric hindrance by poly(ethylene glycol) chain. The influence of the poly(ethylene glycol)–chitosan hybrids on complement component C3 activation was investigated by single radial immunodiffusion method. Influence on complement component C3 activation by poly(ethylene glycol)–chitosan hybrids was almost same as chitosan.     

Echinococcus multilocularis: Immunoglobulin and antibody response in C57BL6J mice

Each of 50 male C57BL/6J mice was infected intraperitoneally with 50 cysts of Echinococcus multilocularis. At 2, 4, 6, 8, and 14 weeks after infection, 10 mice were sacrificed, their larval cyst masses weighed, and their sera collected. Each serum sample from uninfected control and infected mice was adsorbed twice with two batches of E. multilocularis antigen conjugated to Sepharose beads. The concentrations of IgG1, IgG2a, IgG2b, IgM, and IgA in unadsorbed and IgG1, IgG2b, and IgM in adsorbed sera were quantified by the radial immunodiffusion technique. Hydatid mice produced increasingly large amounts of IgG1 and IgM; small measurable increases of IgG2b and no significant increases of IgG2a and IgA were observed during the course of infection. During the rapid growth phase of the cysts (6 to 14 weeks) IgG1 antibodies were found to range from 86 to 93% and IgM antibodies from 17 to 33% of the total IgG1 and IgM. However, the actual protein concentrations of IgM antibodies (761 and 1215 mg/dl) were higher than the sum of the protein concentrations of IgG1 and IgG2b antibodies (411 and 779 mg/dl). The significance of the relative concentrations of IgM, IgG1, IgG2a, and IgG2b antibodies is discussed with reference to their effectiveness in antibody-dependent cellular cytotoxicity and complement-mediated lysis in the control of alveolar hydatid disease.     

Plasmodium berghei: I. Immunochemical properties of fractions of a soluble extract

Soluble extracts of Plasmodium berghei were separated into 12 fractions following preparative disc electrophoresis in polyacrylamide gel. One or two protein bands were detected in each fraction by analytical disc electrophoresis. Similarly, one or two precipitinogens were generally detected in each of Fractions 1 through 11 by immunoelectrophoresis and by double immunodiffusion in agar gel, while the unfractionated extract contained 10 precipitinogens. Antisera produced in rabbits against each fraction each contained two or three (sometimes five) antiplasmodial precipitins demonstrable by immunoelectrophoresis. Serial fractions obtained in separate runs were closely similar to each other, although some degree of overlapping sometimes occurred between neighboring fractions. Glycoproteins were detected in all the fractions, but chiefly in Fractions 4 and 12. The bulk of the RNA in the extract was located in Fraction 4, while hemoglobin was usually confined to Fraction 6. The molecular weights of the soluble components of P. berghei range between 8000 and 130,000.     

Precipitin responses of infected mice to exoantigens and cellular antigens of Trypanosoma musculi.

Plasma were collected from mice which had been immunosuppressed with 650 R from a cobalt-60 gamma radiation source and infected with Trypanosoma musculi. Trypanosomes were also collected from immuno-suppressed mice and from nonirradiated, infected animals. Rabbit antiserum was prepared against trypanosomes fron nonirradiated mice and employed in immunodiffusion analyses to detect trypanosome exoantigens (ExAg) in plasma of irradiated, infected mice and cellular antigens (CAg) in extracts of parasites which had been collected from immunosuppressed and nonirradiated hosts. The rabbit antiserum formed at least 3 precipitin lines with plasma from irradiated, infected mice and 8–9 precipitin lines with extracts of parasites which were obtained from immunosuppressed and untreated mice. Two of the precipitin reactions were against mouse plasma antigens (PAg). Lower levels of PAg appeared to be present in extracts of trypanosomes which were isolated from the irradiated mice than in those from nonirradiated animals.Mice synthesized antibodies against 1 ExAg which was demonstrable in immunodiffusion tests by 14 days after T. musculi infection. A single precipitin reaction was also seen after 21 days. One to 2 precipitin lines were formed with ExAg after 42 days of infection. Two to 3 precipitin lines formed between the ExAg and mouse antisera collected 98, 175 and 341 days after injection of the T. musculi.Similar immunodiffusion reactions were detected with CAg present in both the extracts of T. musculi which had been isolated from irradiated and those from nonirradiated mice and the mouse antisera. One to 2 precipitin lines were found between CAg and antisera from mice which had been infected for 14 days. Two precipitating antigen-antibody systems were seen with antisera collected after 21, 42 and 98 days and 2–3 precipitin reactions were formed between CAg and antisera collected from mice 175 and 341 days after infection.Absorption and immunodiffusion analyses conducted with rabbit and mouse antisera indicated parasite ExAg in plasma of irradiated, T. musculi infected mice were also present in preparations of CAg of the trypanosomes. The persistence of antibody and the increase in the numbers of antigen-antibody systems detected by immunodiffusion during the course of the infection may in part be related to the presence of parasites in capillaries of the kidneys long after they cannot be demonstrated in the peripheral blood of the host.     

Detection of IgG and IgE antibodies to Aspergillus fumigatus in human sera by immunogold assay

An immunogold assay (IGA) was developed to detect IgG and IgE antibodies to Aspergillus fumigatus. Sixteen sera from patients with allergic bronchopulmonary aspergillosis (ABPA), aspergilloma, and normal controls were studied. All sera were also evaluated for antibodies against A. fumigatus by biotin-avidin linked enzyme immunosorbent assay (BALISA) and by agar gel double diffusion method. A. fumigatus specific IgG and IgE antibodies could be detected by IGA in all the patients' sera but not in the sera of normal controls. Both IgG and IgE antibodies to A. fumigatus could be demonstrated in all the sera by BALISA and normal controls showed only low levels of these antibodies. There was a positive correlation between the degree of reactivity detected by IGA, the BALISA titer and the precipitins by agar gel diffusion. It can be concluded that IGA is a reliable, sensitive and simple method capable of detecting both IgG and IgE antibodies against A. fumigatus in patient serum.     

Characterization of invertebrate cell lines

Summary The usefulness of four serologic techniques for distinguishing five selected lepidopteran cell lines was evaluated; the techniques included complement fixation, hemagglutination. immunodiffusion, and immunoelectrophoresis. The five selected lepidopteran cell lines represent three taxonomic families of Lepidoptera with one family, Noctuidae, containing two cell lines derived from insects within the same genus. The five cell lines were crossreactive in complement-fixation tests, but the lines were distinguishable at a familic level when two units of antigens were used in the test. Agglutination of goose erythrocytes was not observed with the antigens over a pH range of 5.8 to 7.2 at 4°C or ambient temperature. Immunodiffusion tests demonstrated a common cross-reactive antigen(s), but spurs of partial identity and the presence of extra precipitin bands were indicative that differentiation at a familic level was possible. Immunoelectrophoresis of the cellular antigens also revealed common cross-reactive precipitin arcs, but the number and clarity of arcs in homologous systems was increased such that four of the five cell lines were distinguishable. A basic protein was consistently seen in the homologous system, but it was absent in the heterologous systems. Although these data suggest that immunoelectrophoresis was the best serologic technique for distinguishing the five lepidopteran cell lines, the shortcomings of this approach are also discussed. This research was supported in part by the World Health Organization, The Rockefeller Foundation, and U.S. Public Health Service Grant AI-13727.