Biomedical subjects

Ellen Kraig

Publications and source records attributed to Ellen Kraig.

6 recordsLinked to original sources

PubMed2003

A hierarchy of T cell receptor motifs determines responsiveness to the immunodominant epitope in experimental autoimmune myasthenia gravis.

The predominant murine T lymphocyte population responding to Talpha146-162, the immunodominant epitope in EAMG, expresses the TCRBV 6 gene segment. However, cells expressing other TCRBV gene segments also react with this peptide. In order to more precisely characterize the Talpha146-162-specific TCR repertoire, we isolated CD4high cells from peptide-immunized mice. The majority of CD4high cells utilized an acidic TCR beta chain CDR3 motif regardless of TCRBV gene usage. Analysis of T cell clones demonstrated a fourfold higher avidity of Vbeta6+ than non-Vbeta6 cells for Talpha146-162 indicating that a hierarchy of TCR motifs determines T cell responsiveness in EAMG.

Amino Acid Motifs↗

PubMed2003

Anaerobic regulation of Actinobacillus actinomycetemcomitans leukotoxin transcription is ArcA/FnrA-independent and requires a novel promoter element.

The periodontal pathogen, Actinobacillus actinomycetemcomitans, produces a 116-kDa leukotoxin that appears to help the bacterium evade the innate host immune response. The expression of leukotoxin is induced when cells are grown anaerobically, a condition found in the subgingival crevice. This regulation most likely occurs at the transcriptional stage since the levels of leukotoxin RNA are induced by hypoxic growth. In order to map the leukotoxin promoter element(s) responsible for oxygen regulation, deletion and linker-scanning mutations were cloned into a transcriptional reporter gene plasmid and then tested in A. actinomycetemcomitans grown aerobically or anaerobically. A 35-bp DNA element, at position -36 to -70, was found to be responsible for the repression of leukotoxin synthesis in aerobically grown A. actinomycetemcomitans. The sequence of this oxygen response element (ORE) does not match the consensus binding sites for known DNA binding proteins, not even Fnr or ArcA which play major roles in oxygen regulation in other bacteria. However, since sequence analysis alone cannot disprove a role for the Fnr or ArcAB pathways in leukotoxin regulation, the genes for the Fnr and ArcA homologues in A. actinomycetemcomitans were identified, mutated by targeted insertional mutagenesis and assessed for loss of oxygen regulation. Deletion of either fnr or arcA altered the expression of numerous A. actinomycetemcomitans proteins, but leukotoxin expression was still repressed by oxygen. These results, coupled with the promoter mutation analyses, lead to the conclusion that A. actinomycetemcomitans employs a novel pathway in the aerobic/anaerobic regulation of leukotoxin synthesis.

Aggregatibacter actinomycetemcomitans↗

PubMed2003

Evidence of a diverse T cell receptor repertoire for acetylcholine receptor, the autoantigen of myasthenia gravis.

We utilized two methods to look for T cell clonal expansions in myasthenia gravis (MG). We analyzed TCRBV CDR3 length polymorphism (spectratyping) to look for evidence of clonal expansion of CD4 or CD8 T cells directly from peripheral blood of MG patients. No statistically significant differences were found between the diversity of TCR repertoires in MG patients compared to normal control individuals when analyzed as groups. Rare oligoclonal expansions were detected in some individual MG patients but the significance of these findings is unclear. Next, we analyzed a panel of T cell hybridomas from acetylcholine receptor (AChR) immunized, MG-susceptible HLA-DR3 transgenic mice. The epitope specificity, TCRBV gene usage and CDR3 sequences of these hybridomas were highly diverse. We conclude there is only limited evidence for restricted TCR repertoire usage in human MG and suggest this may be due to the inability of HLA-DR molecules to select for restricted TCR recognition of AChR epitopes.

Adult↗

PubMed2002

Split tolerance in a novel transgenic model of autoimmune myasthenia gravis.

Because it is one of the few autoimmune disorders in which the target autoantigen has been definitively identified, myasthenia gravis (MG) provides a unique opportunity for testing basic concepts of immune tolerance. In most MG patients, Abs against the acetylcholine receptors (AChR) at the neuromuscular junction can be readily identified and have been directly shown to cause muscle weakness. T cells have also been implicated and appear to play a role in regulating the pathogenic B cells. A murine MG model, generated by immunizing mice with heterologous AChR from the electric fish Torpedo californica, has been used extensively. In these animals, Abs cross-react with murine AChR; however, the T cells do not. Thus, to study tolerance to AChR, a transgenic mouse model was generated in which the immunodominant Torpedo AChR (T-AChR) alpha subunit is expressed in appropriate tissues. Upon immunization, these mice showed greatly reduced T cell responses to T-AChR and the immunodominant alpha-chain peptide. Limiting dilution assays suggest the likely mechanism of tolerance is deletion or anergy. Despite this tolerance, immunization with intact T-AChR induced anti-AChR Abs, including Abs against the alpha subunit, and the incidence of MG-like symptoms was similar to that of wild-type animals. Furthermore, evidence suggests that this B cell response to the alpha-chain receives help from T cells directed against the other AChR polypeptides (beta, gamma, or delta). This model offers a novel opportunity to elucidate mechanisms of tolerance regulation to muscle AChR and to clarify the role of T cells in MG.

Animals↗

PubMed2002

Immunological memory and late onset autoimmunity.

This review will address a paradox that has long fascinated scientists studying the effects of aging on the immune system. Although it has been clearly documented that B and T lymphocytes lose the ability to respond to antigenic or mitogenic stimulation with age, it has nonetheless been noted that the frequency of autoreactive antibodies is higher in older individuals. Given that the majority of the age-associated defects in immune regulation target the naïve T and B lymphocyte subsets, it has been presumed that this increase in antibodies specific for self antigens was due to changes in the B cell repertoire and/or to differences in the mechanisms responsible for generating immune tolerance in primary responses. However, in this review, we will address an alternative possibility that memory immune responses, first generated when the individual was young, may play a critical role in the appearance of serum autoantibodies by reactivation later in life (recall memory). It has recently been shown, in several different systems, that memory immunity can be maintained over the lifetime of the animal. Thus, memory B cells which are self-reactive may be harbored within an organism as it ages and the potential exists that they become re-activated at a later time, resulting in a vigorous autoreactive recall response. This may occur preferentially in older individuals due to several factors, including deficiencies in immune tolerance with age, progressive age-associated loss of tissue integrity yielding neo-self antigens, and possible re-exposure to an infectious agent which induces an autoimmune memory response through molecular mimicry. Thus, we propose that some of the autoantibodies seen in elderly patients and in older animals may have been produced by memory lymphocytes originally generated against antigens encountered during one's youth, but maintained in a tolerant (non reactive) state until a subsequent triggering event occurs. Possible implications of this model will be discussed.

Aging↗

PubMedDate not supplied

Recall immune memory: a new tool for generating late onset autoimmune myasthenia gravis.

Most patients with autoimmune myasthenia gravis (MG) produce autoantibodies against their muscle acetylcholine receptors (AChR), causing debilitating muscle weakness. Approximately 60% of MG patients first exhibit myasthenic symptoms after the age of 40. Yet, in the C57BL/6 mouse model of MG, older mice are resistant to induction of myasthenia gravis. To understand the immunological basis for this resistance, the effects of age on the B-cell responses to AChR from Torpedo californica, the inducing antigen, were addressed. As expected, the primary B-cell response was lower in 20-month-old mice than in 2-month-old mice; the isotype profile was not altered by age. When mice were re-immunized, the anti-T-AChR titers increased in both young and old animals, suggesting that a memory response was elicited. Importantly, memory B-cells activated in young animals were largely resistant to the age-associated loss of immune function and the recall memory response was vigorous. Furthermore, the antibodies produced in re-stimulated older mice were functional, as evidenced by the appearance of MG symptoms in some of these animals. Thus, by eliciting a recall memory response, the first examples of late onset MG in mice have been generated. By analogy, late onset MG in humans may be due to re-activation of B-cell responses initiated at a younger age.

Age of Onset↗