References

Ross, The pathogenesis of atherosclerosis a perspective for the 1990s, Nature 362 801 (1993). 2. D. Steinberg and J.L. Witztum, Lipoproteins and atherogenesis current concepts, J.A.M.A. 264 3041 (1990). 3. G.K. Hansson, L. Jonasson, P.S. Seifert, and S. Stemme, Immune mechanisms in atherosclerosis, Arteriosclerosis. 9 567 (1989). 4. P. Libby and G.K. Hansson, Involvement of the immune system in human atherogenesis current knowledge and unanswered questions. Lab. Invest. 64 5 (1991). 5. G....

Introduction

A general homeostatic mechanism that protects cells and entire organisms from the deleterious effects of environmental stresses is termed the heat shock response. Its product, heat shock proteins (hsps), play major roles in many cellular processes that affect host-parasite interactions. These proteins possess unique properties and have been implicated also in chronic degenerative diseases, infectious diseases, immunology, and cancer. The ability of pathogens to initiate infection, invade...

Conclusions

As detailed in this chapter, hsp expression in arterial endothelium can be induced or augmented by various types of stress, such as hypercholesterolemia, heat treatment, exposure to oxygen radicals or cytokines, ischemia, hemodynamic overload, surgical stress, and infections. Surface exposure of hsp 60 epitope(s) also emerged on endothelial cells. Thus, specific antibodies and T lymphocytes against hsp 60 65 preexisting in circulation could react with surface-exposed hsp 60 components or cross...

CELISA for Detection of Anti Hsp 65 Antibodies

Nunc Immunoplates (Gibco, Paisley, Scotland) with 96 flat-bottom wells were coated with hsp 65 by incubation of a 10 jag ml solution in bicarbonate buffer (pH 9.6) for 2 hr at 37 C. One hundred microliter aliquots of twofold serum dilutions from 1 25 to 1 800 were added in duplicate to the wells and incubated for 1 hr at 37 C. ELISA was performed using incubation for 1 hr at 37 C of a 100-jil volume of peroxidase-conjugated goat-antihuman IgG (diluted 1 4000) (Kirkegaard and Perry Laboratories,...

Development Of A Working Model For The Role Of Hsps In Transplant Immunity

This working model considers four phases in the process of T-cell-induced stress response and the recruitment and activation of hsp-reactive lymphocytes during transplant rejection. Transplant rejection is inititiated by graft-infiltrating alloreactive T cells which through direct cytotoxic effects and indirectly through cytokine release and inflammatory cell recruitment and activation induce allograft injury and tissue stress (phase 1). Stressed target cells in the transplanted tissue respond...

Genetic Location of Hsp 60 Similarity to Nod Idd5 Susceptibility Region

The genomic location of human hsp 60 has also attracted considerable attention. The sequencing and cloning of this gene by Gupta's group have been followed by gene mapping. Using a biotinylation method, we have identified several apparent pseudogenes of the human hsp 60 in metaphase preparation of the human chromosomes (58). Five binding sites of the human hsp 60 gene were located, including one at chromosome 2q32, which is the human equivalent of Table 3 T-Cell Epitope of GAD 65 in NOD Mice...

Hsp 60 Autoimmunity Can Amplify Or Restrain Inflammation

My colleagues and I are beginning to investigate the immunology of natural autoimmunity to hsp 60 by constructing transgenic mice with hsp 60 hyper-expressed under the regulation of different promoters. We are also studying the effects of natural autoimmunity to hsp 60 on the immune response to bacterial antigens. These studies are not yet completed, but it appears that natural autoimmunity to hsp 60 can indeed modify significantly both autoimmune diseases and immune responses to foreign...

A TLymphocyte Activities in Sarcoidosis

Despite the systemic nature of the disease, activated T cells are observed only in the affected organs, whereas T cells of the peripheral blood remain quiescent or may even exhibit decreased reactivity when compared with peripheral blood mononuclear cells (PBMCs) of healthy individuals. The latter phenom enon has given rise to the designation peripheral anergy (43,44), but it can be ascribed to a compartmentalization of lymphocytes and monocytes to affected tissues (28,35,41,44), and it thus...

Hierarchy Of Immunogenic Human Tumor Antigens The Unique The Shared And The Oncogenic

Although the emerging data on CTL epitopes of human melanomas indicate that human tumor antigens may be shared, rumors of the demise of unique antigens on human cancers may be premature (apologies to Mark Twain). The studies carried out so far leave open a significant possibility that they are uncovering an unrepresentative sample of the repertoire (as discussed in Section III). The existence of shared antigens is not in question, but the absence of unique antigens is. After all, shared or...

Functional Mimicry

The essential biological function of hsp 60 molecular chaperons in protein folding and assembly is achieved through the interaction of its peptide-binding domain with target polypeptides. This peptide-binding ability of molecular chaperons resembles a peptide-binding function of MHC classes I and II molecules involved in antigen presentation. After antigen processing in the cell by ATP-dependent proteosomes, the MCH-peptide interaction occurs with those polypeptides, selected from a broad range...

Hspinduced Tlymphocyte Propagation From Heart Transplant Biopsies Correlates With Cellular Rejection

The first study (28) dealt with the incubation of heart transplant biopsies with soluble Mycobacterium tuberculosis extracts (MTEs), a source of hsp recognized by human T cells. Studies on 299 biopsies from 89 heart transplant patients demonstrated that MTE can induce lymphocyte propagation from heart transplant biopsies, especially during rejection. A highly significant correlation was seen between MTE and Il-2-induced lymphocyte growth, and an accelerated growth was seen for cultures...

Correlation Between Hsp Induction And Reduction Of Myocardial Necrosis

Since many of the aforementioned studies have demonstrated that heat shock treatment results in the induction of hsp 72 and a reduction of infarct size after subsequent ischemia and reperfusion, Hutter and colleagues tested the hypothesis that the degree of protection from ischemic injury in heat-shocked animals correlates with the degree of previous hsp 72 induction (34). Rats were pre-treated with 40, 41, or 42 C of whole-body hyperthermia followed by 24 hr recovery. Control rats were...

Stress Proteins As Targets Of Immune Responses

Two characteristics of stress proteins greatly increase the likelihood that these proteins may become autoantigens involved either in initiating or perpetuating autoimmune diseases. First is their great phylogenetic conservation across major species barriers from prokaryotes to mammals. The second is their immuno-dominance that is, their ability to evoke strong immune responses during the course of infections with a large number of infectious agents (detailed elsewhere in this book). Thus,...

Tl And Hsps

There are several studies in which y 8 T cells reactive with syngeneic T cells have been proposed to recognize a complex between a class lb gene product and a peptide derived from a stress protein. A y 8 T-cell hybridoma has been derived from C57BL 6 mice which recognizes syngeneic cells (14). The T-cell receptor expressed by the hybridoma is known to recognize the protein product of the T22b gene. It has been proposed that T22 is complexed with a self-peptide derived from a stress protein. The...

Hsp Expression In The Arterial Intima

Berberian et al. (10) have shown high levels of hsp 70 in human atherosclerotic lesions, particularly in the central portions of atheroma and at necrotic core sites, correlating in intensity with the severity of the lesion. Hsp 70 was also demonstrated to increase arterial cell survival, suggesting that these proteins are associated with a protective mechanism in normal and diseased arteries (11). It is, however, not clear if hsp 70, like hsp 60, is involved in atherogenesis. We studied hsp 60...

A 64kDa AntigenGAD

Most attention has focused on a 64-kDa antigen that was shown through immu-noprecipitation of islet cell extracts with diabetic sera to be a common target of a humoral response (69). A component of the 64-kDa antigen was finally demonstrated to be GAD 65 (70), the major islet isoform of the gamma amino-butyric acid (GABA) shunt enzyme, which is also expressed in several other tissues, including brain, stomach, testis, and, in humans, in the islet a and S cells in addition to the p cell (71)....

Analysis of the Frequencies of STCLs Stimulated with the Selected Peptides in the Ocular Arthritic and Mucocutaneous

Further analysis of BD into ocular, arthritic, and mucocutaneous types revealed that lymphocytes from patients with the ocular type yielded the highest frequency of stimulation with the four hsp peptides (five of six patients), especially peptides 111-125 and 311-326. In the arthritic type of BD, lymphocytes from four of the six patients responded to these peptides, and there was no obvious difference in stimulation between the four peptides, although the highest frequency was elicited by...

Acknowledgments

R. van der Zee and Dr. W. van Eden for providing the monoclonal antibodies LK-1 and LK-2 Dr. J.D.A. van Embden for providing the recombinant mycobacterial hsp 65 and Dr. M. de Boer for providing the B7-24 monoclonal antibody. The production and distribution of recombinant hsp 65 are supported by the UNDP World Bank WHO special programme for research and training in tropical diseases. Part of the present work was supported by a grant of the National Fund for Scientific...

Viihsp 60specific T Cells In Atherosclerosis

In healthy humans, circulating T cells reacting with mycobacterial hsp 65 have been observed to cross react with shared epitopes of human hsps (57). All animals used in our studies showed circulating T cells reacting with hsp 65 at significantly higher levels than with ovalbumin, which was used as a nonrelevant control antigen (6,12). As described, hsps showed very high phylogenetic sequence homology, not only between bacterial genera but also throughout all higher forms of life. Life-long...

Mechanism Of The y8 Tcell Responses To Hsps

The precise mechanisms of any of the antigen responses of y8 T cells await further clarification. In the case of the polyclonal antigen responses, it seems clear that the y8 T-cell reactivity has different requirements than those of either conventional antigen or superantigen responses of ap T cells. These include requirements on the TCR, the stimulators (antigens), and the accessory cells. The polyclonal response of murine yS T cells to hsp 60 is still the best-studied example. The response...

Hsp

Hsp 70 homologues occur in all the known groups of organism, that is, the archebacteria, the eubacteria, and the eukaryotes, and are the most highly conserved proteins found so far. For example, the eukaryotic hsp 70 homologues are all between 50 and 98 identical in aminoacyl sequence. This high conservation makes these proteins ideal for studies of evolutionary relationships a recent comparison (35) concludes that the archebacteria are more closely related to the gram-positive bacteria than to...

Intracellular Expression of Stress Proteins

It is well recognized that many stimuli other than heat, such as drugs, hormones, certain prostaglandins, mitogens, and cytokines, can induce the synthesis of stress proteins in human cells. For example, synthesis of hsp 90 is dramatically, albeit transiently, increased in T cells following stimulation with mitogens (4), and the stress response may be part of a negative-feedback loop that downregulates the expression of genes for interleukin-1 p (IL-lp) and perhaps other cytokines (5). Because...

Hsps And Fungal Diseases

It has been proposed that the heat shock response plays a fundamental role in infectious processes during host invasion. In addition to bacterial infections (e.g., Mycobacterium, Shigella, and Salmonella), parasites (e.g., Leishmania, Trypanosoma, Plasmodium, Giardia, and Schistosoma), hsps have been detected in many pathogenic fungi (e.g., Aspergillus fumigatus, Fonsecae pedrosoi, and Histoplasma capsulatum), when, at the time of infection, the organisms adapt to temperature increases....

Pathogenesis

The pathogenesis of Crohn's disease remains unknown. Research focuses at the possible initiating events, the proinflammatory mediators, and deficient down-regulatory mechanisms of intestinal inflammation. The role of the mucosal immune system is thought to be of major importance in this respect (3). There is accumulating evidence that a microbial factor may be involved in the initiation or amplification of the intestinal inflammation in Crohn's disease. In an animal model, nonsteroidal...

Immune Responses to Mycobacterial Hsp 60 in Diabetes

The potential role of the heat shock protein hsp 60 in the development of diabetes in NOD mice was first postulated by Elias et al. (6,7). This hypothesis was based on the detection of T-cell and antibody responses against a 62-kDa molecule cross reactive with a mycobacterial 65-kDa heat shock protein. These hsp 65 cross-reactive T-cell and antibody responses developed coincidently with the progression of insulitis and were followed by induction of anti-insulin antibodies and anti-idiotypic...

Hsp 60 or Chaperonins

These proteins were originally identified in several laboratories in the 1970s as host-encoded proteins required for the morphogenesis of several bacteriophages in E. coli. An operon called groE was identified encoding two protein subunits, one around 60 kDa called GroEL and the other around 10 kDa called GroES (reviewed in Ref. 52). Both proteins are essential for the growth of E. coli at all temperatures, and it was known that GroEL is immunologically similar to a dominant immunogen of...

Cytokines in IDDM Direct Effectors or Promoters of Cell Mediated Immunity or Autoantigen Expression

Several lines of evidence suggest that cytokines have an important role in the destruction of the p cells in IDDM either by direct cytotoxicity or by a cytokine imbalance promoting other effector mechanisms. Interleukin-lp (IL-ip) is currently the main candidate for a direct effector (33). The relative sensitivity of islet cells to the cytotoxic effects of IL-ip and the high molar excess of the natural receptor antagonist required to inhibit the cytokine have been demonstrated in vitro....

Tcell Responses

Murine Repertoire Following infection with M. tuberculosis or M. bovis BCG, T-cell proliferation interferon gamma (INF-y) and other cytokine responses to hsp 65 and hsp 16 have been reported (32,33). The magnitude of hsp responses was similar to that found for other protein antigens. The hsp 65 epitope specificity of T cells from draining lymph nodes of primed mice was examined in respect to seven selected peptides (34). Immunodominant as sell as cryptic epitopes had been...

The Contradictions Of Hsp 60 Autoimmunity

The conservation of stress proteins throughout evolution may be explained by their performance of essential functions and by the dependence of this performance on particular structures encoded in unique amino acid sequences to maintain the functions of stress proteins, evolution, during its meandering course, has had to preserve their key sequences. Other chapters in this book discuss the functions and structures of stress proteins. Whatever these functions may be, the conservation of the amino...

BCD8 T Cells

Cloned murine cytotoxic cells of CD8 phenotype, restricted by H-2 class I molecules, were reported to be cytolytic against target cells expressing both mycobacterial and autologous hsp 65 molecules (61). The stimulatory autologous macrophages needed preincubation with INF-y or cytomegaloviral infection, which presumably led to enhanced expression of the target hsp 60 molecule. In another context, CD8 T cells purified from the spleens of M. vaccae-primed mice have been found active as suppressor...

D18kDa aCrystallin

This protein is a member of a family of low molecular weight hsp a-crystallins which has only about 30 of sequence conservation. The 18-kDa protein initially found in M. leprae, responds to heat shock in M. simiae (habana) serovar 1 (25). It has also been found to be present in the M. avium complex (containing two genes) and several other species of mycobacteria but not in M. tuberculosis (26). The failure to find homologous gene in M. tuberculosis is of concern in view of cross reactivity,...

Hsps and Cross Reactivity Between Nonself and Self

Numerous studies have reported autoimmune recognition of hsp (for summary, see ref. 6), but the relation between priming of hsp-specific lymphocytes by nonself-hsp and subsequent response to cross-reactive self-hsp epitopes is poorly understood. T cells recognizing epitopes from hsp 60 shared among microbes and humans have been identified in healthy individuals (50,51). By using an in vitro stimulation system with a tryptic peptide digest of mycobacterial hsp 60, CD8 T lymphocytes could be...

Human Visceral Leishmaniasis and the Humoral Response to

In the search for fusion proteins that could be used for diagnosis, MacFarlane et al. (15) used a .gtll library constructed from L. donovani promastigote mRNA (52) and found one clone that was recognized by serum antibodies from an African patient with kala-azar. Sequence analysis of this clone revealed that its 600-bp insert coded for the carboxyterminal region of an antigen related to the 70-kDa heat shock protein family. The full-length sequence of the corresponding gene (1959 nucleotides)...

Cy5 T Cells

Increase of y8 T cells have been reported in normal and athymic mice immunized with M. tuberculosis in incomplete Freund's adjuvant (63,64). yS T cells were also induced by PPD-containing aerosol in resident pulmonary lymphocytes (65). Increase of y5 T cells has also been reported in M. bovis BCG-in-fected mice (66,67). Although M. tuberculosis has been the antigenic stimulus in these murine studies and also for human Vy9 Vy2 T cells, representing a major population in normal individuals (68),...

Hsp 60 As A Pcell Antigen In Iddm

Hsp 60, previously referred to as hsp 65, is the human equivalent of the Escherichia coli GroEL protein and mycobacterial hsp 65 (36). It is a member of the chaperonin family of proteins, whose diverse functions include protein folding and cellular protection (37). The amino acid sequences are highly conserved between species, with greater than 65 homology between the human and mycobacterial forms (36). The hsp 65 of lower organisms is considered to be immunodominant. Following infection or...

Stress Proteins as Molecular Chaperones

University of Warwick, Coventry, England All cells contain groups of highly conserved proteins that increase rapidly in concentration when the cells are exposed to environmental stresses. The most studied stress is a temperature 5-10 C higher than that optimal for the growth of the cell being studied, and thus these proteins are often called heat shock proteins (or hsps). However, the same set, or in some cases a subset, of these proteins increases in amount when cells are subjected to a whole...

Causal Involvement Of Hsp 60 In Atherogenesis

Evidence for a causal involvement of hsp65 in atherogenesis is provided by our animal models induced by immunization or feeding a cholesterol-rich diet (6). As mentioned earlier in this chapter, we have immunized rabbits one or more times with various antigens, with or without adjuvants. The antigens and adjuvants, respectively, included human and rabbit atherosclerotic lesion proteins, ovalbumin, recombinant mycobacterial hsp 65, Freund's complete and incomplete adjuvants, and two hsp-free...

Proposed Model For Stress Protein Induction In

We propose the following model for the mechanisms involved in the induction of a stress response in DMD (Fig. 3) and postulate that ROS play a key role in Ca dependent activation or proteases Ca dependent activation of proteases Figure 3 Proposed model for selective stress protein induction in DMD. See text for details. the induction of hsp 70 and oxidation-specific stress proteins. Hsp 70 may be directly upregulated by ROS which lead, in human cells, to translocation and binding of HSF-1 to...

The Clonal Selection Paradigm

According to the classic formulation of the clonal selection theory (2), the essence of the immune system and its distinctive feature is its repertoire of antigen receptors. These receptors, which include the T- and B-cell antigen receptors and the antibodies, are produced by somatic genetic recombinations and mutations (3) and so are created anew in each individual. The selective force that determines the frequencies of the various receptors in the repertoire is exerted by the antigens that...

Immune Pathogenesis Of C Trachomatis Infections

Although it is well known that C. trachomatis infections can cause infertility and adverse pregnancy outcome, the mechanism whereby this occurs has only recently begun to be elucidated. In an in vitro fallopian tube organ culture, addition of C. trachomatis induced little damage to the integrity of the fallopian tubes (2). Similarly in vivo, experimental inoculation of C. trachomatis into the fallopian tubes of nonhuman primates resulted in only a transient infection with no long-lasting damage...

Humoral Immune Responses

Humoral immune responses to the C. trachomatis 57-kDa hsp have been associated with female infertility and upper genital tract infections. High-titer antibody to the chlamydial 57-kDa hsp was most prevalent in women who were infertile owing to fallopian tube occlusion (8,9) and in women with pelvic inflammatory disease who had an ectopic pregnancy (10). The relation between a humoral immune response to the chlamydial 57-kDa hsp and tubal infertility due to a C. trachomatis infection has been...

Ppd As A Carrier Molecule In Animals Sensitized With

Bacille Calmette-Guerin (BCG, attenuated Mycobacterium tuberculosis var. bovis, used as a vaccine against tuberculosis) has been used in the past to non-specifically enhance the immune response to antigens coinjected with it (9). In addition to this, Lachmann and coworkers originally reported (10) that mice first primed with BCG and then immunized in the absence of adjuvants with B-cell epitopes conjugated to purified protein derivative (PPD, tuberculin, currently used for diagnostic skin...

Brief Review Of The Clinical Features Of Lyme Disease

Lyme disease (LD) is a multisystem inflammatory disease caused by B. burgdorferi. As reviewed (29,30), the clinical features of the disease can be divided into three categories early localized (31), early disseminated (32,33), and chronic or late disease (34,35). The immunology of Lyme disease has been reviewed previously (36,37). A. Early Localized LD (Formerly Stage I) The pathognomonic skin rash of LD, erythema migrans (EM), seen in about 50-70 of patients, usually begins as an erythematous...

The Immunological Homunculus

The paradoxes of hsp 60 autoimmunity can be ascribed to a principal misconception of the classic clonal selection theory the behavior of the immune system is not controlled solely by chance contacts with antigens it is no less constrained by the internal organization of the immune system (24). This organization reflects the evolutionary experience of the species. For example, the capacity of the receptor repertoire to interact with particular antigens is dictated by various elements encoded in...

Iiistress Proteins During Development Of The Nervous System

Increasing evidence suggests that stress proteins may play an important role during nervous system development. This was demonstrated in both drosophila and mammalian systems (2-5). In drosophila, (2,3), mRNA for heat shock proteins (hsps) 23 and 26 were constitutively expressed during particular stages of larval development, mainly in neurocytes and thoracic ganglion cells. In rat embryos (4,5), severe heat shock resulted in marked deformities in the forebrain and eyes. Less severe heat shock...

Heat Shock Proteins in the NOD Mouse

Immunostimulation Reduces the Incidence of IDDM in the NOD Mouse Several studies have established that the incidence of IDDM in the NOD mouse can be reduced by exposure to mycobacterial preparations including complete Freund's adjuvant (killed Mycobacterium tuberculosis in oil emulsion) (41-43) and the bacille Calmette-Guerin vaccine (BCG, live attenuated Mycobacterium bovis) (44). The mode of action is unclear, but hypotheses have included nonspecific immune stimulation, activation of...

Hsp Function In Muscular Diseases

We propose that the induction of stress proteins deserves protective functions in muscle diseases. This hypothesis is based on observations from many different groups and in various models, both in vitro and in vivo, for a protective function of stress proteins, as well as from direct evidence showing that mdx mice fed an iron-free diet display simultaneously less muscle necrosis and less hsp expression (L. Bornman et al., submitted). Potential mechanisms for protection by stress proteins in...

Beneficial Effects of Hsps

Beneficial effects of hsps based on their chaperone functions have been reported. Evidence for autoprotection of the infected cell by elevated hsp synthesis during infection has been obtained with human cells (62-67). Increased expression and supply of the inducible hsp 72 by astrocytes during scrapie may contribute to neuronal homeostasis by preventing the deposition of neurotoxic amyloid proteins. This could retard damage of the central nervous system, at least in the initial phase of disease...

Vsp Expression In Tissues Monocytes And Macrophages In Sarcoidosis

Tissue biopsies, collected for diagnostic purposes and prior to any treatment, from patients with various interstitial lung diseases (ILDs), including sarcoidosis, were immunolabeled using SP-specific murine monoclonal antibodies (MAbs). The ML30 MAb was produced against M. leprae (95) and recognizes M. lepra SP 65 as well as a cross-reactive sequence in human SP 60 (reviewed in ref. 20). Two other SP 60-specific MAb, LK-1 and LK-2, were produced against human SP 60 (96). LK-2 cross reacts with...

Clinical Relevance Of Hsp Overexpression

As discussed above, hsp 90 was significantly elevated in those patients with SLE who had severely active disease compared with those patients who had inactive or mildly active disease using the UCH Middlesex scoring system (12). Using a more sophisticated system, the British Isles Lupus Assessment Group's (BILAG) system for analyzing disease activity, disease activity was assessed in eight different disease categories or organ systems. Hsp levels were correlated with active or inactive disease...

In Vitro Fertilization Outcome And Immune Responses To

Women who are infertile owing to blockage of the fallopian tubes are prime candidates for IVF. The possible effect on IVF outcome of being sensitized to the chlamydia 57-kDa hsp was, therefore, investigated in a prospective study (16). Endocervical samples were obtained from 216 women at the time of oocyte retrieval and tested by ELISA for immunoglobulin A (IgA) antibodies reactive with the recombinant C. trachomatis 57-kDa hsp. Among the 68 women whose IVF cycle resulted in a term birth, only...

Overview Of The Structure And Function Of The Stress Proteins

The mammalian stress proteins often are divided into two groups based on their classic mode of induction the heat shock proteins (hsps) and the glucose-regulated proteins (grps) (reviewed in refs. 5 and 6). In general, both the hsps and grps are identified by their apparent molecular mass as determined by SDS-PAGE (Table 1). Members of the grp family were first observed to exhibit increased expression in cells starved of glucose. Subsequently, the grps were shown to also undergo high-level...

Autoimmunity In Chronic Lyme Disease

It is not clear if persistence of the organism is necessary for ongoing disease, since antibiotic therapy may not be as effective in late disease as it usually is at an early stage (38). Indeed, the presence of B. burgdorferi at local sites of inflammation is difficult to demonstrate (37). These findings are compatible with the premise that an autoimmune mechanism may be active in the pathogenesis of chronic Lyme disease. Although an immune response to heat shock proteins in patients with Lyme...