Projects

Project 1 - David Topham
Project 2 - Andrea Sant

Project 1: David Topham

The overall goal of the clinical core is to provide samples from carefully characterized subjects after influenza infection or vaccination, to support the conduct of studies under projects 1 (Determinants of heterosubtypic immunity against influenza); 2 (Specificity of the CD4 T cell immune response to influenza); and 3 (Effector functions of influenza-specific T cells induced by immunization and infection). The clinical core provides for collection of appropriately timed samples from individuals receiving routine vaccination or after natural infection and from individuals participating in clinical trials supported by other mechanisms, and provides sample processing, storage, and tracking.

In support of this mission, four clinical protocols have been generated and these protocols are summarized briefly.

Studies of the immune response to influenza are usually limited to assessment of secondary infections, because primary exposure to influenza occurs early in life, typically before the age of nine years. Assessment of these responses is complicated by lack of knowledge of the specific influenza virus responsible for earlier exposures, and the timing of previous exposures. These problems is compounded as persons age and have experienced multiple infections. One approach to obtaining samples from primary or near primary infection is to perform prospective surveillance in populations of children with relatively high attack rates. Therefore, this study involves prospective surveillance conducted in families where at least one child is under the age of four years.

Although the family study is ideal for assessing primary, or near primary infections, a disadvantage of this approach is that limited numbers of samples, and limited amounts of blood, are feasible to be obtained in the context of a family surveillance study. As a result, we are supplementing these measurements by obtaining serial measurements of immunity in college students and other young adults who are likely to remain in contact with the study center for at least 3 influenza seasons. The primary objective of this surveillance activity is to obtain relatively larger amounts of cells, including pre-influenza season samples.

The purpose of the third protocol is to provide cells and plasma from normal donors to develop and validate additional assays of influenza immunity. These assays will then be used in studies to assess immune responses to vaccination and infection in children, adults, and the elderly. Healthy donors undergo medical screening and rapid assessment of hematocrit.

The last study is built on previous studies that demonstrated the ability of previous vaccination with a clade 0 H5N1 vaccine to prime for responses to a single dose of clade 1 H5N1 vaccine administered 8 years later. That study was limited by a relatively small study sample size, and the lack of concurrent testing of a vaccine naïve group. The primary objectives of this study are to describe the kinetics of the cellular immune response to subvirion H5N1 vaccination in primed and unprimed individuals, to determine the effects of priming on the repertoire and phenotypes of B and T cells generated in response to vaccination, and to generate hypotheses regarding the relationship between the peptide specificity of CD4 T cells prior to vaccination and subsequent humoral and cellular responses. This will be a randomized, subject and laboratory blinded assessment of the cellular immune response to A/Indonesia/5/05 vaccine in two populations: (1) healthy adults who have previously received an H5 vaccine in a DMID sponsored study; and (2) healthy adults with no previous H5 vaccination and who are not at risk for H5 exposures. Subjects will be randomized to either a low dose (15 mcg) or high dose (90 mcg) of vaccine, and peripheral blood mononuclear cells will be obtained before and at several time points after each dose of vaccine, for assessment of peptide-specific responses of CD4 T cells, cytokine and cell marker phenotypes of responding helper CD4 T cells, numbers of antigen-specific antibody secreting cells by B cell ELISPOT, the presence and quantity of antigen specific memory B cells, and flow cytometric correlates of B cell and antibody responses.

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Project 2: Andrea Sant

The defining feature of the immune system is its ability to distinguish self from non-self, a function mediated by antigen-specific T lymphocytes. T cell receptors can only recognize antigens derived from pathogens or transformed cells if these antigens if the derived peptide fragments of pathogenic proteins combine with Major Histocompatibility Complex (MHC) molecules. The assembly of the antigenic peptide-MHC complex takes place in intracellular compartments, by a series of molecular events collectively referred to as "MHC-restricted antigen presentation". The research in my laboratory centers around the molecular events that regulate MHC class II-restricted antigen presentation and CD4 T cell activation in vivo. Our long term goal is to make connections between the mechanisms involved in peptide acquisition by class II molecules and those aspects of immunology that critically depend on the specific peptides presented by the class II molecule.

Our laboratory has a long-standing interest in the mechanisms involved in establishment of immunodominance in CD4 T cell responses. During immune responses pathogens or to protein antigens, T lymphocytes only respond to a limited number of peptide epitopes contained in the immunogens. These peptides are termed "immunodominant". Our experiments seek to understand the elements in vivo that dictate the narrowed selection of specificities in CD4 T cells during protective immune response. Most recently, we have begun to explore the hierarchy of CD4 T cells responses to influenza virus, a human pathogen that poses unique challenges for vaccine design because of its high degree of genetic variability and because of the recent threat of avian influenza. The research area in our laboratory focuses on understanding the molecular and cellular events that shape and refine the repertoire of CD4 T cells that are specific for influenza virus, using both animal models and analyses of human immune responses. We also seek to understand the specificity of human CD4 responses to influenza virus and vaccines during the primary response to influenza infection,, during the establishment of long term memory and during challenge with vaccines or live viruses. Or research on influenza has the long-term goal of rational vaccine design to develop vaccines that promote heterosubtypic immunity in the CD4 T cell compartment, by developing strategies to focus the CD4 T cells towards the most biologically active and genetically conserved epitopes. We are developing strategies to dissect MHC class II restricted presentation of influenza antigens by B cells in order to understand the mechanisms that control recruitment of CD4 T cell help by B cell during influenza infection, a requisite event for the production of high affinity neutralizing antibodies to influenza virus.

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Get in Touch

For general questions, call:
Donna Neu
(585) 276-5621

Email: Donna Neu

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