Your browser doesn't support javascript.
loading
Show: 20 | 50 | 100
Results 1 - 20 de 41
Filter
Add more filters










Publication year range
1.
Genes Immun ; 14(8): 504-11, 2013 Dec.
Article in English | MEDLINE | ID: mdl-24067789

ABSTRACT

Toll-like receptors recognize several components of Mycobacterium tuberculosis, the main causative agent of tuberculosis. The signaling pathways leading to activation of the immune response require the MyD88 and TIRAP genes. The hypothesis that polymorphic variants of these genes influenced resistance to pulmonary tuberculosis was tested by a case-control study (400 cases and 400 controls). Heterozygosity at the polymorphic sites MyD88 rs6853 (alleles: A, G) or TIRAP rs8177374 (S180L) (alleles: C, T) is associated with resistance to pulmonary tuberculosis (P: 7.8 × 10(-8) and 2 × 10(-6), respectively). Double heterozygosity confers higher protection levels (P: 10(-14) to 2 × 10(-16)). The logistic regression model displayed that the double homozygous genotype GG/TT predisposes to the disease (odds ratio (OR): 5.78) and the AG/TT genotype combination neutralizes the protective activity exerted by AG (OR: 3.05). The same model showed that the risk of developing the disease increases with age from 31-40 years to 71-80 years (OR: 1.32-13.59).


Subject(s)
Disease Resistance/genetics , Membrane Glycoproteins/genetics , Myeloid Differentiation Factor 88/genetics , Polymorphism, Single Nucleotide , Receptors, Interleukin-1/genetics , Tuberculosis, Pulmonary/genetics , Adult , Age Factors , Aged , Amino Acid Sequence , Case-Control Studies , Female , Heterozygote , Humans , Male , Membrane Glycoproteins/chemistry , Middle Aged , Models, Genetic , Molecular Sequence Data , Myeloid Differentiation Factor 88/chemistry , Receptors, Interleukin-1/chemistry , Tuberculosis, Pulmonary/immunology
2.
Genes Immun ; 10(7): 641-6, 2009 Oct.
Article in English | MEDLINE | ID: mdl-19536151

ABSTRACT

Patients (305 patients with pulmonary tuberculosis) and controls (290 household genetically unrelated contacts) were tested by polymerase chain reaction (PCR) for polymorphisms in the intron 15 and the 5' untranslated region of the gene coding for the a3 isoform of the human ATPase gene. Diagnosis of pulmonary tuberculosis was based on chest radiography and sputum smear examination and confirmed by PCR and bacteriological tests. Alleles (two at each site) segregated in the form of four haplotype pairs: 13, 14 (very rare), 23, and 24. The 13/24 (double heterozygous) patients were protected against tuberculosis (OR: 0.15; P: 10(-8); CI: 0.08-0.3). The 13/13 vs 13/24 and 23/23 vs 23/24 (double homozygous) patients were susceptible to the disease (OR. 5.8; P: 6 x 10(-7); CI: 2.8-11.9; OR: 4.5; P: 5 x 10(-7); CI: 2.5-8.4, respectively).


Subject(s)
Gene Frequency/genetics , Genetic Predisposition to Disease , Protein Isoforms/genetics , Tuberculosis, Pulmonary/genetics , Vacuolar Proton-Translocating ATPases/genetics , Adult , Alleles , Female , Genotype , Haplotypes/genetics , Heterozygote , Homozygote , Humans , Male , Middle Aged
3.
J Appl Microbiol ; 106(6): 2041-7, 2009 Jun.
Article in English | MEDLINE | ID: mdl-19298512

ABSTRACT

AIMS: To ascertain whether in Brucella abortus-infected water buffalo herds, the number of newly infected animals could be reduced by culling superspreaders (the animals secreting > or =10(4) CFU per ml of milk). METHODS AND RESULTS: The number of B. abortus present in the milk (CFU per ml) from 500 water buffaloes was measured by the culture. Each animal was tested three times, at one month intervals. The presence or the absence of B. abortus in each milk sample was confirmed by PCR. A majority of infected animals shed the pathogen at a low level (< or =10(3) CFU ml(-1)). However, a few infected individuals (superspreaders) shed large numbers of B. abortus (> or =10(4) CFU ml(-1)). Quantitative PCR of B. abortus positive milk samples gave comparable results to culture. Culling of the superspreaders was sufficient to arrest the spread of infection. SIGNIFICANCE AND IMPACT OF THE STUDY: The approach described here can reduce significantly the cost of controlling brucellosis. Culture and quantitative PCR tests identify superspreaders and, compared with the serological tests in use to detect brucellosis, provide also a more accurate estimate of the disease incidence.


Subject(s)
Brucella abortus/isolation & purification , Brucellosis, Bovine/diagnosis , Buffaloes/microbiology , Milk/microbiology , Animals , Antibodies, Bacterial/blood , Brucella abortus/genetics , Brucella abortus/immunology , Brucellosis, Bovine/microbiology , Brucellosis, Bovine/prevention & control , Buffaloes/immunology , Cattle , DNA, Bacterial/genetics , Female , Flow Cytometry , Polymerase Chain Reaction/veterinary
4.
Immunogenetics ; 60(3-4): 157-65, 2008 Apr.
Article in English | MEDLINE | ID: mdl-18330558

ABSTRACT

A case-control study established that the haplotype pair HYA/HYA at the MBL (mannose binding lectin) locus of water buffalo is associated with resistance to Brucella abortus infection (P < 10(-7)) and the haplotype pairs LYD/LYD with susceptibility to the same pathogen (P < 10(-7)). The subjects included in the present study were tested twice-at a 1-month interval-for the presence of anti-B. abortus antibodies in the serum by agglutination, complement fixation and flow cytometry. Cases (335 subjects) included animals consistently positive to all these tests; controls (335 subjects) comprised animals exposed yet negative by the same tests. The serum from genetically resistant subjects displayed in vitro significantly higher antibacterial activity compared to the serum from genetically susceptible subjects, lending biological significance to the results from the association study. Inhibition of the antibacterial activity following heat treatment of the serum, addition of specific MBL inhibitors (EDTA, mannose, N-acetyl-D: -glucosamine) or anti-human MBL antiserum provide convincing evidence that the antibacterial activity present in the serum results from the interaction between MBL and B. abortus. A replication study (comprising 100 cases and 100 controls) confirmed the results from the original study.


Subject(s)
Brucella abortus , Brucellosis/veterinary , Buffaloes/genetics , Buffaloes/immunology , Haplotypes , Mannose-Binding Lectin/genetics , Alleles , Animals , Blood Bactericidal Activity , Brucellosis/genetics , Brucellosis/immunology , Case-Control Studies , Mannose-Binding Lectin/immunology
5.
Clin Microbiol Infect ; 12(3): 248-53, 2006 Mar.
Article in English | MEDLINE | ID: mdl-16451412

ABSTRACT

A bacteriophage lytic for Escherichia coli O157:H7 was isolated from bovine manure. Following in-vivo selection, the phage acquired the capacity to persist in the circulatory system of mice for at least 38 days. When mice were infected experimentally with E. coli O157:H7 (10(7) CFU/mouse), simultaneous injection of the mice with phage (10(8) PFU/mouse) cleared E. coli O157:H7 from the mice within 48 h.


Subject(s)
Coliphages/isolation & purification , Escherichia coli Infections/microbiology , Escherichia coli O157/virology , Manure/virology , Viremia , Animals , Coliphages/physiology , Escherichia coli Infections/prevention & control , Escherichia coli O157/isolation & purification , Female , Lysogeny , Mice , Mice, Inbred BALB C
6.
J Microbiol Methods ; 55(1): 109-19, 2003 Oct.
Article in English | MEDLINE | ID: mdl-14500002

ABSTRACT

Phage-resistant mutants have been isolated from Streptococcus thermophilus. Selection was carried out using anti-phage antibodies or Hoechst 33258-labelled phages. Two mutants out of eight tested displayed reduced acidifying capacity. Selection of the bacteria that extruded more rapidly the fluorochrome 5-6-carboxyfluorescein diacetate (CFDA) restored the acidifying capacity of these two mutants to the level of the parental strains. Mutants displaying phage resistance and good acidifying capacity were obtained in 4-5 days. New phages that are able to overcome the protection mechanisms of the existing bacteria arise continually in the dairy environment. The procedures described here permit to replace promptly the starter culture susceptible to newly emerged phages with a resistant one.


Subject(s)
Lysogeny , Mutation , Streptococcus Phages/genetics , Streptococcus/genetics , Adsorption , Antibodies, Viral/immunology , Bisbenzimidazole/metabolism , Polymerase Chain Reaction , Streptococcus Phages/immunology
7.
Cytometry ; 37(3): 246, 1999 Nov 01.
Article in English | MEDLINE | ID: mdl-10520207
8.
J Clin Microbiol ; 36(3): 802-6, 1998 Mar.
Article in English | MEDLINE | ID: mdl-9508316

ABSTRACT

Two flow cytometric assays are described herein. The single cytometric test (SCT) detects antibodies to either Brucella abortus or Staphylococcus aureus in the serum or milk of a cow or water buffalo. The double cytometric test (DCT) detects both anti-B. abortus and anti-S. aureus antibodies concurrently. In the SCT, the sample to be tested is incubated in succession with the antigen (either B. abortus or S. aureus) and the proper secondary antiserum (fluorescein isothiocyanate-labelled rabbit anti-cow immunoglobulin antiserum or rabbit anti-water buffalo immunoglobulin antiserum). In the DCT, the sample to be tested is incubated first with B. abortus and S. aureus antigens and then with the secondary antiserum. The B. abortus antigen used in the DCT is covalently bound to 3-microm-diameter latex particles. The difference in size between B. abortus and S. aureus permits the establishment of whether the antibodies are directed against one, the other, or both antigens. When compared to the complement fixation test, the SCT and DCT each show a specificity and a sensitivity of 100%. The SCT has been used previously to detect anti-S. aureus antibodies. Here its use is extended to the detection of anti-B. abortus antibodies. The DCT is described here for the first time. The DCT appears to be useful for large-scale brucellosis eradication programs. It offers the possibility of using one test to identify animals that are serologically positive for both B. abortus and S. aureus.


Subject(s)
Antibodies, Bacterial/analysis , Brucella abortus/immunology , Flow Cytometry , Milk/immunology , Staphylococcus aureus/immunology , Animals , Antibodies, Bacterial/blood , Brucellosis/diagnosis , Brucellosis/veterinary , Brucellosis, Bovine/diagnosis , Buffaloes , Cattle , Cattle Diseases/diagnosis , Fluorescent Antibody Technique , Sensitivity and Specificity , Staphylococcal Infections/diagnosis , Staphylococcal Infections/veterinary
9.
Cytometry ; 29(3): 261-6, 1997 Nov 01.
Article in English | MEDLINE | ID: mdl-9389443

ABSTRACT

The nuclear DNA content of seven mouse laboratory strains has been measured by flow cytometry. The differences observed between strains as well as those between sexes within the strain were all statistically significant. The highest DNA content (approximately 6.4 pg/female nucleus) was found in the Balb/c strain; the lowest (approximately 5.7 pg/male nucleus) in the C3H/he strain. The difference between sexes varied from 1.6% (in CD-1 mice) to 6.3% (in nude mice). The interest of these results is twofold. First, the mouse can now be used to study the adaptive significance of genome size variation, so far studied only in plants. Second, DNA content analysis can become a quick method for mouse strain identification.


Subject(s)
DNA/analysis , Flow Cytometry/methods , Mice, Inbred Strains/genetics , Animals , Cell Nucleus/chemistry , Female , Male , Mice
10.
Nat Toxins ; 5(1): 14-9, 1997.
Article in English | MEDLINE | ID: mdl-9086454

ABSTRACT

Seiridin (SE), one of the main phytotoxins produced in vitro by Seiridium species pathogenic to cypress, was oxidized and the corresponding ketone derivative covalently linked to bovine serum albumin (BSA). The conjugate (SE-BSA) was used to prepare an antiserum to SE. The antibodies were absorbed with BSA and their specificity was assayed by ELISA and flow cytometry against SE, iso-seiridin (ISE), a structural isomer of SE, and some derivatives of these two metabolites. The antibodies tested in a competitive indirect ELISA did not show any binding activity to SE, ISE and their derivatives. The cytometry test, instead, was successful. SE-BSA and SE showed the highest binding activity with the antibodies. SE derivatives having a shift on the adjacent carbon, oxidation, or acetylation of the hydroxy group of the heptyl side chain at C-4 or conversion of the gamma-lactone in the corresponding planar furane ring reacted less than SE. The 2'-dansylhydrazoneSE and the 3,4-dihydroSE having a bulky group attached to the heptyl side chain and a saturated lactone ring, respectively, showed a weak reactivity. SE derivatives in which the gamma-lactone ring was destroyed and ISE derivatives presenting the shift of the hydroxy group at C-3' and another structural modification had no binding activity.


Subject(s)
Furans/analysis , Mitosporic Fungi/chemistry , Mycoses/etiology , Mycotoxins/analysis , Plant Diseases/microbiology , Trees/microbiology , Animals , Antibody Specificity , Enzyme-Linked Immunosorbent Assay/methods , Flow Cytometry/methods , Furans/immunology , Immunoglobulin G/analysis , Mitosporic Fungi/pathogenicity , Mycotoxins/immunology , Rabbits , Sensitivity and Specificity , Serum Albumin, Bovine/analysis
11.
J Virol Methods ; 69(1-2): 137-45, 1997 Dec.
Article in English | MEDLINE | ID: mdl-9504759

ABSTRACT

The simultaneous detection is described of cucumber mosaic virus (CMV), potato virus Y (PVY) and tomato mosaic virus (ToMV) by flow cytometry. Extracts from leaves of healthy and CMV or PVY infected plants were incubated with latex particles, each with a diameter of 3 microm. Extracts from ToMV infected or uninfected plants, however, were incubated with particles, each with a diameter of 6 microm. Beads were washed and incubated in succession with primary and secondary antibodies, the latter labeled with phycoerythrin (PE) or fluorescein (FITC). CMV and PVY were distinguished on the basis of the fluorescence emitted by FITC and PE; ToMV was distinguished from CMV and PVY on the basis of the different diameter (6 microm) of the particles on which it was adsorbed. The three viruses were detected also by another approach. Latex particles with a diameter of 3, 6 and 10 microm were separately sensitized with antibodies specific for CMV, PVY and ToMV. An equal number of sensitized particles was mixed and incubated with the plant extracts containing the three viruses and then with anti-CMV, anti-PVY and anti-ToMV antibodies labeled with FITC. The study describes also a virus purification method based on the use of antibody coated latex particles. The method is simple technically and applicable to the purification of large as well as minute amounts of different viruses (CMV, PVY and ToMV).


Subject(s)
Cucumovirus/isolation & purification , Flow Cytometry , Plant Leaves/virology , Plant Viruses/isolation & purification , Potyvirus/isolation & purification , Tobamovirus/isolation & purification , Antibodies, Viral/immunology , Fluorescein , Microspheres , Phycoerythrin , Plant Extracts , Reproducibility of Results , Sensitivity and Specificity
12.
Res Vet Sci ; 60(2): 179-81, 1996 Mar.
Article in English | MEDLINE | ID: mdl-8685542

ABSTRACT

An assay has been developed to detect antibodies to Staphylococcus aureus in water buffalo milk by flow cytometry. The method was the protein A-deficient strain Wood 46 of S aureus incubated with milk samples and fluorescein-labelled rabbit anti-water buffalo antiserum. The assay can detect antibodies when the pathogen is not detectable by bacterial tests and can determine the antibody titre directly on undiluted samples.


Subject(s)
Antibodies, Bacterial/analysis , Buffaloes , Milk/immunology , Milk/microbiology , Staphylococcus aureus/immunology , Animals , Female , Flow Cytometry/methods , Rabbits , Sensitivity and Specificity
14.
J Immunol Methods ; 154(2): 211-6, 1992 Oct 02.
Article in English | MEDLINE | ID: mdl-1401954

ABSTRACT

A procedure is described for the selection of mouse Lyt2- mutant cells. The procedure is based upon repeated cycles of selection with rat monoclonal antibodies to the Lyt2 antigen and magnetic particles coated with goat anti-mouse IgG. Stable Lyt2- mutant clones were derived from cells previously mutagenized with X rays and, at a lower frequency, also from non-mutagenized cells. The procedure can be used to rescue Lyt2- cells when the ratio of Lyt2+:Lyt2- cells is 10(4):1.


Subject(s)
Antigens, Ly/genetics , Cell Separation/methods , T-Lymphocyte Subsets/cytology , Animals , Antigens, Ly/analysis , Immunologic Techniques , In Vitro Techniques , Lymphocyte Depletion , Magnetics , Mice , Mutation
15.
Hybridoma ; 9(2): 149-55, 1990 Apr.
Article in English | MEDLINE | ID: mdl-2347597

ABSTRACT

Monoclonal antibodies specific for an allotypic marker of goat IgG2 were used to select goat-mouse hybrid cells secreting goat IgG2. Four of these hybrid cell clones continued to synthesize goat IgG2 (5-15 micrograms/ml) for over eight months. They will be used to study goat IgG gene regulation.


Subject(s)
Goats/immunology , Hybridomas/immunology , Immunoglobulin G/metabolism , Animals , Cell Fusion , Cell Line , Female , Lymphocytes , Mice , Multiple Myeloma
16.
Anim Genet ; 21(2): 171-80, 1990.
Article in English | MEDLINE | ID: mdl-2386313

ABSTRACT

Five allotypic determinants controlled by independent genes have been identified in goat. Of these determinants, four have been detected with alloimmune antisera and one with monoclonal antibodies. The specificities A1, C1 and D1 are lipoproteins; B1 is possibly an alpha 2 macroglobulin and E1 and IgG2. The specificity B1 is not expressed until the age of 3-4 months. The gene controlling the specificity E1 is present at about the same frequency (0.38-0.41) in goat, sheep, cattle and water buffalo. Stable hybridomas secreting goat IgG2 have been obtained by the fusion of goat peripheral lymphocytes with mouse myeloma cells.


Subject(s)
Goats/genetics , Hybridomas/metabolism , Immunoglobulin Allotypes/genetics , Immunoglobulin G/biosynthesis , Mice/genetics , Animals , Antibodies, Monoclonal , Cell Fusion , Gene Expression , Goats/immunology , Immunoglobulin Allotypes/analysis , Recombination, Genetic
17.
Hybridoma ; 8(3): 315-21, 1989 Jun.
Article in English | MEDLINE | ID: mdl-2473027

ABSTRACT

A monoclonal antibody against goat immunoglobulins recognizes an allotypic determinant (A1) which is common to goat, sheep, cattle and water buffalo. The frequency of the corresponding gene (A') is about the same in all four species, indicating the existence of a polymorphism that remained stable over a period of about 18-20 million years.


Subject(s)
Antibodies, Monoclonal , Isoantigens/genetics , Ruminants/immunology , Animals , Buffaloes , Cattle , Epitopes/genetics , Goats , Hybridomas/immunology , Mice , Phylogeny , Polymorphism, Genetic , Ruminants/genetics , Sheep , Species Specificity
18.
J Dairy Res ; 54(4): 471-7, 1987 Nov.
Article in English | MEDLINE | ID: mdl-3693628

ABSTRACT

In order to standardize a radioimmunoassay of milk progesterone as a routine method for confirmation of oestrus and diagnosis of pregnancy in water buffalo, monoclonal antibodies against progesterone were produced. Hybridomas were prepared by fusing spleen cells from a Balb/c mouse immunized with progesterone 11 alpha-hemisuccinate-bovine serum albumin conjugate with the mouse myeloma cell line NS-1. Thirty wells out of 94 secreted anti-progesterone antibodies. Of the ten independent hybridomas derived, one (AF65) was suitable for the quantification of milk progesterone by radioimmunoassay. The tracer used in the assay was progesterone-11 alpha-hemisuccinate [( 2-125I]iodohistamine). The sensitivity of the assay was 50 pg/tube. The mean progesterone concentration at oestrus was 0.8 +/- 0.2 ng/ml increasing to 8.5 +/- 0.8 ng/ml 24 d later in pregnant animals.


Subject(s)
Antibodies, Monoclonal , Breeding/methods , Buffaloes/physiology , Estrus Detection/methods , Milk/analysis , Pregnancy Tests/veterinary , Progesterone/analysis , Animals , Female , Pregnancy , Pregnancy Tests/methods , Radioimmunoassay
20.
Int Arch Allergy Appl Immunol ; 64(1): 115-7, 1981.
Article in English | MEDLINE | ID: mdl-7450900

ABSTRACT

The paper describes a rhesus monkey serum antigen (D1) which is controlled by a dominant gene (D1), independent from those controlling A1, B1 and C1, which are three low-density lipoprotein markers.


Subject(s)
Antigens/immunology , Macaca mulatta/immunology , Macaca/immunology , Absorption , Animals , Antigens/genetics , Chromatography, Affinity , Chromatography, Gel , Chromatography, Ion Exchange , Female , Genes, Dominant , Immune Sera/pharmacology , Immunoelectrophoresis , Immunoglobulin Allotypes , Lipoproteins, LDL/genetics , Macaca mulatta/genetics , Male
SELECTION OF CITATIONS
SEARCH DETAIL
...