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1.
Sci Rep ; 9(1): 4834, 2019 03 18.
Article in English | MEDLINE | ID: mdl-30886198

ABSTRACT

The complexity of immune responses limits the usefulness of univariate methods in answering complex immunology questions. To demonstrate the utility of a multivariate approach, we employ such approach to compare T cells of African green monkeys (AGMs) and rhesus macaques (RMs). Among the most prominent distinguishing features we found were lower CD3 and higher CD28 surface expression in AGMs compared to RMs. After in vitro stimulation, a larger proportion of AGM T cells secreted cytokines, especially those producing more than one cytokine (i.e. multifunctional cells). To find out whether multifunctional responses associate with protection in other species, we compared T cells of cynomolgus macaques (CMs) infected with wild-type Simian Immunodeficiency Virus (SIV) to those of CMs infected (vaccinated) with a replication-defective virus. Wild-type SIV infection in macaques leads to simian Acquired Immunodeficiency Syndrome (AIDS), which does not happen in animals previously vaccinated with a replication-defective virus. Interestingly, after in vitro stimulation, multifunctional cells were more abundant among T cells of vaccinated CMs. Our results propose T-cell multifunctionality as a potentially useful marker of immunity, although additional verification is needed. Finally, we hope our multivariate model and its associated validation methods will inform future studies in the field of immunology.


Subject(s)
Immunologic Techniques/methods , SAIDS Vaccines/immunology , Simian Acquired Immunodeficiency Syndrome/immunology , T-Lymphocyte Subsets/immunology , T-Lymphocytes/immunology , Animals , CD28 Antigens/metabolism , CD3 Complex/metabolism , Chlorocebus aethiops/immunology , Chlorocebus aethiops/virology , Cytokines/immunology , Cytokines/metabolism , Immunogenicity, Vaccine , Lymphocyte Count , Macaca mulatta/immunology , Macaca mulatta/virology , SAIDS Vaccines/administration & dosage , Simian Acquired Immunodeficiency Syndrome/blood , Simian Acquired Immunodeficiency Syndrome/virology , Simian Immunodeficiency Virus/genetics , Simian Immunodeficiency Virus/immunology , Simian Immunodeficiency Virus/pathogenicity , Species Specificity , T-Lymphocyte Subsets/metabolism , T-Lymphocytes/metabolism , Virus Replication/genetics , Virus Replication/immunology
2.
Diagn Microbiol Infect Dis ; 63(3): 296-301, 2009 Mar.
Article in English | MEDLINE | ID: mdl-19117714

ABSTRACT

Methicillin-resistant Staphylococcus aureus is a significant problem in healthcare settings around the world. To effectively track different strains, we used a number of genetic procedures. In this study, 252 clinical isolates from the Milwaukee, WI, area were analyzed by the multiplex polymerase chain reaction (PCR) method to determine which of the described types were present. Most strains were categorized into one of the known groups, although types III and IIIA strains were not found. An unusual variant was found in 17% of the samples. This variant (v1) contains an 800-base pair amplimer that was created from a unique configuration of 2 primers from different sets in the multiplex PCR procedure. Although superficially similar to a type I pattern, sequencing of the amplimer demonstrated that it is identical to a sequence from the USA300 strain, a type IV strain. It is suggested that this variant may have resulted from acquisition of an IS431 element near the dcs region. There is potential for misidentification of strains because of slight genetic alterations such as this one. Statistical analysis using a chi(2) contingency table demonstrated that amoxicillin/clavulanic acid resistance was significantly lower in types I and IA when compared with the other types, whereas erythromycin resistance was significantly lower in type I when compared with other types. However, there was not a significant difference for any of the new variants.


Subject(s)
DNA Fingerprinting/methods , DNA, Bacterial/genetics , Methicillin-Resistant Staphylococcus aureus/drug effects , Methicillin-Resistant Staphylococcus aureus/isolation & purification , Staphylococcal Infections/epidemiology , Staphylococcal Infections/microbiology , Anti-Bacterial Agents/pharmacology , Bacterial Typing Techniques , DNA Transposable Elements , Genotype , Humans , Microbial Sensitivity Tests , Polymerase Chain Reaction/methods , Sequence Analysis, DNA , Wisconsin/epidemiology
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