Search PubMed⌕ Search

SEARCH · Search PubMed

Results for “IFA”

Search indexed PubMed citations on genomics, clinical trials, systematic reviews and public health. Explore titles, authors and supplied subject terms, then open the PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 955 records · Page 53Linked to original sources

[Evaluation of an ELISA technique for the serodiagnosis of legionnaires' disease].

The presence of anti-Legionella antibodies was studied comparatively in 81 sera with a commercially available ELISA kit (containing serogroups 1 to 6 as antigens) and the indirect immunofluorescence assay (IFA) with serogroup 1. The values obtained with the ELISA method were converted into immunofluorescence titers by using standard sera. Results of both methods were concordant in 83% of the sera when a cutoff value of 1/128 was used, and in 88% of the sera when the cutoff value was 1/128 for IFA and 1/256 for ELISA. In 6 other patients infected by serogroups other than 1, antibodies were detected in 5 cases with ELISA and in none with IFA.

Enzyme-Linked Immunosorbent Assay↗

Serosurvey for Cowdria ruminantium, Coxiella burnetii, and Spotted fever group rickettsiae in ostriches (Struthio camelus) from Zimbabwe.

Sera from 216 ostriches on nine farms around Zimbabwe were examined by indirect fluorescent antibody (IFA) testing for the presence of antibodies reactive with Cowdria ruminantium, Coxiella burnetii, and Rickettsia africae, a spotted fever group rickettsia. Although no reactive antibodies could be detected to C. ruminantium or C. burnetii, 51/216 (35%) sera reacted with R. africae. The seroprevalence in ostriches from the south of Zimbabwe was significantly higher than in birds from the north (P < 0.01). Immunoblots of four sera positive by IFA (> 1/160) showed antibodies reactive with antigens of R. africae that also were recognized by pooled sera from mice inoculated with the organism. No reactive antibodies could be detected in six sera negative by IFA.

Animals↗

Evaluation of fetal infection and abortion in pregnant ponies experimentally infected with Ehrlichia risticii.

Fetal infectivity of Ehrlichia risticii was investigated in 19 ponies that were E risticii negative on the basis of results of an indirect fluorescent antibody (IFA) test. Thirteen pregnant ponies were infected by IV administration of E risticii between 90 and 180 days of gestation. Six pregnant ponies served as noninfected controls. Each infected pony had clinical signs of equine monocytic ehrlichiosis, was confirmed to be ehrlichemic, and developed an IFA titer to E risticii. Two infected ponies became recumbent, were unresponsive to supportive care, and were euthanatized. After recovery from clinical illness, the remaining ponies were observed throughout gestation for reproductive abnormalities. On abortion, each fetus was necropsied and tissue specimens from the liver, bone marrow, spleen, colon, and mesenteric lymph nodes were inoculated into canine monocyte cell cultures. Six infected ponies aborted at a mean 217 days of gestation, which was between postinoculation days 65 and 111. Five fetuses were recovered for evaluation, and E risticii was isolated from 4 of them. All 5 fetuses recovered had similar histologic finding, including enterocolitis, periportal hepatitis, and lymphoid hyperplasia with necrosis of the mesenteric lymph nodes and spleen. All 5 fetuses tested negative for IgG to E risticii, although 3 had low IgM titer to E risticii. The remaining 5 infected ponies had normal parturition. Presuckle IFA titer to E risticii was measured in 4 of the term foals, and results for 3 were positive. Two foals from infected ponies were monitored for 6 months and daily gain in body weight was comparable to that of a control foal. None of the control ponies became ill or seroconverted during the clinical illness phase, and none aborted throughout gestation Two control ponies seroconverted to E risticii 6 weeks before parturition. Results of this study indicate that E ristcii is a primary abortifacient under experimental conditions.

Abortion, Veterinary↗

Antibody-mediated basophil accumulations in cutaneous hypersensitivity reactions of guinea pigs.

Cutaneous basophil hypersensitivity (CBH) was studied in guinea pigs by using simplified histologic techniques. Animals immunized with oxazolone or picryl conjugates of keyhole limpet hemocyanin (KLH) emulsified with complete (CFA) or incomplete Freund's adjunvant (IFA) were found to have hapten-specific cutaneous basophil reactions when skin tested at 1 week with oxazolone or picryl chloride contant painting or intradermal injection of oxazolone or picryl-conjugated human serum albumin, respectively. Thus, hapten-specific cutaneous basophil reactions were present in guinea pigs immunized with CFA for classical delayed hypersensitivity, and in animals immunized with IFA for Jones-Mote reactions. Hapten-specific 24-hr cutaneous basophil reactions were passively transferred with immune serum from donors sensitized with conjugates of oxazolone or picryl-KLH in CFA or IFA, and with serum from oxazolone contact-sensitized animals as well. As little as 0.5 ml sera obtained from donors 1 week after immunization could systemically transfer cutaneous basophil reactions. It is likely that hapten-specific cutaneous basophil reactions are mediated by small quantities of serum antibodies. We conclude that antibody-mediated cutaneous basophil reactions may be distinctive hypersensitivity responses that can be distinguished from classical anaphylactic, Arthus, and delayed hypersensitivities. It is suggested that CBH reactions are heterogeneous and that antibody products of B lymphocytes, and factors probably derived from T lymphocytes, play a role in basophil accumulations at cutaneous hypersensitivity reactions.

Animals↗

Surveillance of legionellosis and Legionella bacteria in the built environment in Singapore.

During the period 1986 to 1996, a total of 258 sporadic cases of community-acquired legionellosis was reported, giving a mean annual morbidity rate of 0.9 per 100,000 population. Majority of the cases (91.5%) was serologically diagnosed by the indirect fluorescent antibody (IFA) test against Legionella pneumophila based on single sera with a titre of > or = 1:1024. The others were confirmed by a four-fold increase in IFA titre to a minimum of 1:128 (7.4%) and by culture (1.1%). Cases were reported predominantly among males, ethnic Indians, the elderly and those with concurrent medical conditions. The overall case-fatality rate was 14.7%, with the rate significantly higher among those aged > or = 45 years (21%), smokers (20.4%) and those with concurrent medical conditions (22.5%). Seroepidemiological studies among the healthy population based on an IFA titre of > or = 1:32 showed a prevalence of between 10.3% and 21.9%. Of the various occupational groups investigated, high seroprevalence rates were found in foreign construction workers (52.6%) and occupants of a building investigated for an unusual building-related illness (76.1%). Environmental surveillance of Legionella bacteria showed that 36% of cooling towers, 15% to 19% of decorative fountains and waterfalls and 2% of spa pools were positive. The majority (85%) of environmental isolates was identified as Legionella pneumophila: 54.8% of serogroup 1 and 5% of Pontiac subtype.

Adolescent↗

Purification of intact nuclear lamina and identification of novel laminlike proteins in Raji, a cell line devoid of lamins A and C.

Research on the structure of the nuclear lamina and the nuclear matrix of cells devoid of lamins A and C has been hampered by the fact that intact residual nuclear structures are difficult to isolate from such cells. In this paper, we show that some extraction parameters, such as buffer composition and the nature of the detergent used to remove nuclear membranes, are critical for achieving isolation of whole nuclear residual structures from the lymphoblastic cell line Raji, used as a model for cells without lamins A and C. Electron microscopic analysis shows that the nuclear lamina of Raji cells is formed by a network of intermediate-size filaments interrupted with circular discontinuities. Both lamins B1 and B2, and lamin D/E, are present in this structure. In addition, a group of 45-kDa proteins or intermediate filament protein--reacting proteins (IFA-RPs), located uniquely in the lamina, were found to exhibit the same immunological and chemical characteristics as lamins. Although they behave like nuclear lamins, microsequencing analysis of the IFA-RPs has revealed no homology with known lamins. These IFA-RPs may contribute to the formation of the nuclear lamina filament network in the absence of lamins A and C.

Blotting, Western↗

Antigenic similarity between Hammondia hammondi and Toxoplasma gondii tachyzoites.

Hammondia hammondi is an obligate heteroxenous intestinal coccidian of cats, sharing many characteristics with Toxoplasma gondii. The tachyzoite stage antigens of T. gondii and H. hammondi were studied by immunofluorescence assays (IFA) and western blotting (WB) techniques to demonstrate antigenic similarities. Five monoclonal antibodies (MAbs), anti-T. gondii antigens, P22, P23, P30, P35, and P43, and mice polyclonal anti-H. hammondi serum were investigated. Antigens of H. hammondi were recognized by anti-P30 MAb both in IFA and in WB and by anti-P22 and anti-P35 MAbs only in IFA. Polyclonal anti-H. hammondi serum revealed many common antigens between the 2 parasites (30, 32, 35, 66, and 90 kDa). The differences of host parasite relationship between these 2 coccidians lead us to suggest that many of these antigens with similar molecular weights are not the same, but homologous, molecules or that they are not the only factors involved in these differences.

Animals↗

[Plasmapheresis in multimodality therapy of alveolitis].

Fifty one patients with alveolitis of different genesis (idiopathic fibrotic alveolitis (IFA) in 15, extrinsic allergic alveolitis (EAA) in 20, fibrotic alveolitis (FA) in 10, and eosinophilic pneumonias (EP) in 6), receiving multimodality therapy had a session of plasmapheresis (PA) including 3-4 procedures at a week interval. The session was repeated in 4 patients. The earliest results in all patients appeared as better well-being, improved parameters of blood and external respiratory function and half the patients had positive X-ray changes. The best results were achieved by patients with EP, the worst by those with IFA. Following 2 years, remission retained in a third of patients, recrudescence was noted in two thirds. Five patients with IFA and EAA died. In most patients PA was started late when they had developed recrudescence and respiratory failure. Ineffective therapy made the authors think of PA. The latter improved all the patients' condition and yielded early positive changes in many parameters. PA may have a positive impact in the late period.

Alveolitis, Extrinsic Allergic↗

Prevalence of and risk factors for fecal shedding of Cryptosporidium parvum oocysts in horses.

OBJECTIVE: To determine prevalence of and risk factors for fecal shedding of Cryptosporidium parvum oocysts among 3 populations of horses. DESIGN: Cross-sectional study. ANIMALS: 152 horses participating in the 1996 Texas State 4-H Horse Show, 144 horses examined by the veterinary teaching hospital, and 70 broodmares and their 10- to 21-day-old foals. PROCEDURE: Information on signalment and potential risk factors for fecal shedding of oocysts was obtained. Fecal samples were evaluated for oocysts by means of acid-fast (AF) staining, immunofluorescence assay (IFA), and, for selected samples, flow cytometry (FC). RESULTS: Results of the 3 diagnostic tests were significantly different. The best agreement was between results of the IFA and FC; AF staining and FC were more sensitive than the IFA, but AF staining was less specific. Fecal samples from 13 horses were classified as positive for oocysts. Risk factors for fecal shedding of oocysts included residence on 2 of 4 breeding farms involved in the study, age < 6 months, and history of diarrhea during the preceding 30 days. A municipal water source was implicated as a risk but could not be verified because of insufficient data. CLINICAL IMPLICATIONS: Mature horses and exposure to cattle did not appear to be important sources of cryptosporidial infection for foals. Overall prevalence of C parvum infection among these horses was low, and C parvum infection appeared to be associated with particular farms, rather than an endemic opportunistic infection in horses.

Animals↗

Detection of Rickettsia species with seroreactivity to Rickettsia tillamookensis and Rickettsia rickettsii subsp. californica in domestic dogs in Northern California.

Rickettsia species phylotype G022 (G022) and Rickettsia tillamookensis both maintain a low infection prevalence in Ixodes pacificus ticks and share a close phylogenetic history with pathogenic Rickettsia species. The transmission mechanisms and pathogenic potential of I. pacificus-borne Rickettsia are poorly understood. The low prevalence of G022 and R. tillamookensis suggests that vertebrate species are utilized for maintenance, though identification of vertebrate amplifier and reservoir hosts utilized by Rickettsia is limited. To evaluate the molecular occurrence of G022 and R. tillamookensis and the sentinel utility of domestic dogs, a molecular survey was conducted. Buffy coat and blood plasma samples from 175 domestic dogs collected from Humboldt and Sonoma counties in northern California were screened for rickettsial DNA and anti-rickettsial antibodies, respectively. Dogs were screened for antibodies against Rickettsia spp. and tested for seroreactivity against R. tillamookensis, Rickettsia rickettsii subsp. californica, and Rickettsia rhipicephali. Because G022 has not yet been isolated, this Rickettsia species was omitted from the serological survey. Of the 175 dogs tested, three (1.71%; 95% CI: 0.58%-4.90%) were seropositive for Rickettsia spp. by indirect immunofluorescence antibody assay (IFA); two dogs (1.14%; 95% CI: 0.31%-4.10%) demonstrated seroreactivity by enzyme-linked immunosorbent assay (ELISA), suggesting previous exposure to Rickettsia spp. Among dogs from Sonoma County, 2.88% (3/104; 95% CI: 1.0%-8.14%) of plasma samples were seropositive for Rickettsia spp. by IFA, and 1.92% (2/104; 95% CI: 0.53%-6.74%) demonstrated seroreactivity by ELISA, while all the dogs from Humboldt County were seronegative for both methods. Although the ELISA results were not considered species-specific, two plasma samples demonstrated the greatest seroreactivity against R. tillamookensis and R. rickettsii subsp. californica, respectively, suggesting previous exposure to Rickettsia spp. with epitope similarity to R. tillamookensis and R. rickettsii subsp. californica, respectively. No DNA was detected for G022 or R. tillamookensis by quantitative real-time PCR, with a limit of detection of approximately 3.6 target copies per reaction for G022 and 1 genomic copy per 2&#xa0;&#x3bc;l of template DNA per reaction for R. tillamookensis.

Animals↗

Role of serology in the diagnosis of Lyme disease.

Numerous concerns regarding the potential for misdiagnosis of Lyme disease using commercial assays have been voiced by the US Food and Drug Administration (FDA). We attempted to clarify the clinical value of serologic testing for Lyme disease using the results of commonly marketed assays for detecting antibody to Borrelia burgdorferi, the organism that causes Lyme disease. We reviewed published studies on B burgdorferi test performance published through 1998, package insert labeling from FDA-cleared test kits for B burgdorferi, and Lyme Disease Survey Set LY-A from the College of American Pathologists. We assessed the sensitivity and specificity of commercial serologic tests (enzyme-linked immunosorbent assay [ELISA], immunofluorescence antibody [IFA], and immunodot) for diagnosis of Lyme disease. To reduce this risk of misdiagnosis, it is important that clinicians understand the performance characteristics and limitations of these tests. These tests, in common use in clinical or commercial laboratories, should be used only to support a clinical diagnosis of Lyme disease, not as the primary basis for making diagnostic or treatment decisions. Serologic testing is not useful early in the course of Lyme disease because of the low sensitivity of tests in early disease. Serologic testing may be more useful in later disease, at which time sensitivity and specificity of the test are improved. Positive or equivocal results on an ELISA, IFA, or immunodot assay requires supplemental testing with a Western blot assay. A negative result on the Western blot or ELISA indicates that there is no serologic evidence of infection by B burgdorferi at the time the sample was drawn.

Antibodies, Bacterial↗

Absence of serological evidence for foamy virus infection in patients with amyotrophic lateral sclerosis.

Foamy virus (FV) infection has been implicated in the pathogenesis of sporadic motor neuron disease (MND) by means of serological assays. To confirm these results we tested serum and cerebrospinal fluid (CSF) samples from 23 cases of clinically verified non-familial MND and 11 cases of suspected non-familial MND for the presence of FV infection as determined by Western blot (WB) and indirect immunofluorescence assay (IFA). Using the same tests we also screened sera from 87 healthy chimpanzees for the presence of FV antibodies. None of the human samples in question tested positive. However, the testing revealed that 84 of 87 chimpanzees (96.6%) were seropositive for FV, indicating that combined WB and IFA are suitable methods for the serodiagnosis of FV infection. Given these results an association of FV infection and sporadic MND is highly improbable. Furthermore a suggested therapeutic trial with anti-retroviral drugs appears unjustified.

Adult↗

Immune responses to inactivated vaccine in people naturally infected with hantaviruses.

An inactivated Hantaan virus vaccine for hemorrhagic fever with renal syndrome (HFRS) was given by injection to 15 people who were naturally infected with either Hantaan or Seoul viruses. Immunofluorescent antibody (IFA), reversed passive hemagglutination inhibition (RPHI), hemagglutination inhibition (HI), and neutralization antibody (NA) assays were used to measure the antibody titers of the vaccinated people before and after three doses of vaccine. The results indicated that IFA and RPHI antibody titers were boosted significantly (P < 0.05) after the vaccination. Either Hantaan or Seoul virus could induce two-way cross-reactive neutralization antibody responses in humans. After HTNV vaccine immunization, the NA titers of people with natural infection increased significantly (P < 0.05) to both Hantaan and Seoul viruses, while the relative dominance between these two type responses was still similar to that of natural infection. It is worthwhile to studying the procedure further to inoculate two different virus vaccines for improving the cross-protective effect.

Adolescent↗

In situ indirect fluorescent antibody: a new specific test to detect ongoing chagasic infections.

In situ immunofluorescence assay (ISIFA) was developed for the selection for treatment of chronically infected chagasic patients and their follow-up. Trypomastigotes flash fixed in situ on microscopic slides with 0.025% glutaraldehyde were used as antigen. ISIFA results were compared with complement-mediated lysis (CoML), membrane immunofluorescence (MbIFA), immunofluorescence assay (IFA) with epimastigotes, and xenodiagnosis (XENO). ISIFA was able to distinguish nontreated chagasic patients (geometric mean titer [GMT] = 180) and treatment failures (GMT = 160) from those considered successfully treated (GMT = 25). ISIFA revealed a high sensitivity and titers of 80 or higher detected 98.6% of patients with active infections, even in those with negative XENO, CoML, or MbIFA. Specificity evaluated in 63 sera from other infections, including leishmaniasis and autoimmune diseases, was 98%. IFA used in routine diagnostic procedures exhibited similar results in all groups, irrespective of therapy.

Adult↗

The enhanced antigen-specific production of cytokines induced by pertussis toxin is due to clonal expansion of T cells and not to altered effector functions of long-term memory cells.

Pertussis toxin (PT) has been shown to act as an adjuvant that enhances the production of both Th1 and Th2 cytokines to coinjected protein antigens. It has remained unresolved, however, how PT affects the clonal sizes, long-term effector functions, and Th1/Th2/Th0 differentiation of the T cell responses induced. We have studied the effects of PT on the development of the CD4(+) T cell response to a prototypic antigen, hen eggwhite lysozyme (HEL). HEL injection with incomplete Freund's adjuvant (IFA) resulted in an IFN-gamma(-)/IL-5(+) Th2 recall response. In comparison, co-administration of PT with HEL:IFA enhanced the frequencies of IL-5-producing T cells up to eightfold, and induced the differentiation of high frequencies of IFN-gamma-producing CD4(+) T cells. The results showed that the IFN-gamma and IL-5 produced, originated from clonally expanded Th1 and Th2, but not Th0 cells, and that the effector functions of long-term memory cells were unaffected. Adoptive transfer experiments suggested that PT mediated these effects via activation of APC, not by acting on the T cells directly. The effects of PT on the developing T cell response required the presence of the holotoxin (A- and B-subunit); the individual subunits did not show adjuvant effects. The data suggest that PT enhanced cytokine production by promoting differentiation and vigorous clonal expansion of Th1 and Th2 cells via activation of APC.

Animals↗

Oligodeoxynucleotides containing CpG motifs can induce T cell-dependent arthritis in rats.

OBJECTIVE: To investigate whether CpG oligodeoxynucleotides (ODNs) can induce or accelerate arthritis in rats. METHODS: The CpG-induced response was studied by recording joint inflammation, cell activation in draining lymph nodes, and levels of the acute-phase reactant alpha(1)-acid glycoprotein (AGP) in sera. The role of T cells was investigated by in vivo administration of monoclonal antibodies specific for the T cell receptor alpha/beta (TCRalpha/beta), followed by analysis of cell phenotypes by flow cytometry. RESULTS: One intradermal injection of CpG ODN emulsified with Freund's incomplete adjuvant (IFA) induced arthritis in LEW and LEW.1AV1 rats, while the control ODN sequence without CpG motifs or IFA alone did not trigger disease. The CpG/IFA and control-ODN/IFA injections induced lymphoplasia as well as elevated levels of interleukin-1beta and interferon-gamma messenger RNA in lymph nodes. The arthritis was preceded by elevated levels of AGP in serum. In vivo administration of anti-TCRalpha/beta antibodies after disease induction caused decreased expression of the TCR-CD3 complex on circulating T cells and ameliorated the arthritis. CONCLUSION: We demonstrated that injection with immunostimulatory CpG, both in phosphorothioate-modified and native forms, can induce a T cell-dependent joint-specific inflammation in LEW and LEW.1AV1 rat strains. This arthritis is preceded by signs of activation of the innate immune system. Since unmethylated CG dinucleotides are common in bacterial DNA but rare in mammalian DNA, our results indicate that exposure to bacterial DNA during infection may contribute to arthritis induction by amplifying the innate immune response.

Animals↗

Requirement of dying cells and environmental adjuvants for the induction of autoimmunity.

OBJECTIVE: Cells commonly die without eliciting autoimmunity. However, dying cells are a potential initiating stimulus for systemic lupus erythematosus (SLE). Our goal was to verify whether immune adjuvants influence the autoimmunity induction that ensues following in vivo injection of dying cells. METHODS: Mice were immunized with apoptotic thymocytes in the presence of artificial moieties, such as Freund's incomplete adjuvant (IFA), or natural adjuvants, such as dendritic cells (DCs). Renal involvement and the development of autoantibodies were monitored. RESULTS: Apoptotic cells failed to induce clinical disease or to sustain production of autoantibodies in (NZB x NZW)F(1) mice. In contrast, autoimmunity developed in the presence of IFA or DCs. The characteristics of the adjuvant influenced the array of autoantibodies, the kinetics of their development, and the severity of the disease. DCs were required for induction of anti-beta(2)-glycoprotein I IgG. Adjuvants alone did not elicit disease. CONCLUSION: A "two-hit" signal composed of autoantigens and adjuvants initiates systemic autoimmunity. Moreover, environmental signals at the site of clearance of dead cells shape the features and the severity of the autoimmune disease. Strategies aimed at preventing the accumulation of dying cells and at modulating endogenous adjuvants may be beneficial for the treatment of SLE.

Animals↗

Monoclonal antibodies against plant proteins recognise animal intermediate filaments.

Four monoclonal antibodies were raised against polypeptides present in a high-salt detergent-insoluble fraction from cells of Chlamydomonas reinhardtii. Indirect immunofluorescence microscopy of fibroblasts and epithelial cells grown in culture using these plant antibodies revealed staining arrays identical to those obtained with well characterised antibodies to animal intermediate filaments. Immunofluorescence microscopy of Chlamydomonas with these monoclonal antibodies and a monoclonal antibody that recognises all animal intermediate filaments (anti-IFA) gave a diffuse, patchy cytoplasmic staining pattern. Both the plant antibodies and anti-IFA stained interphase onion root tip cells in a diffuse perinuclear pattern. In metaphase through to telophase, the labelling patterns colocalised with those of microtubules. Labelling of the phragmoplast was also detected but not staining of the preprophase band. On Western blots of various animal cell lines and tissues, all the antibodies labelled known intermediate filament proteins. On Western blots of whole Chlamydomonas proteins, all the antibodies labelled a broad band in the 57,000 Mr range, and three antibodies labelled bands around 66,000 and 140,000 Mr but with variable intensities. On Western blots of whole onion root tip proteins, all the antibodies labelled 50,000 Mr (two to three bands) polypeptides and a diffuse band around 60,000 Mr and three of the antibodies also labelled several polypeptides in the 90,000-200,000 Mr range. The consistent labelling of these different bands by several different monoclonal antibodies recognising animal intermediate filaments makes these polypeptides putative plant intermediate filament proteins.

Antibody Specificity↗