Interplay of antibody and T cell responses in acute myocardial infarction.

This study sought to investigate the interplay between antibody and T cell responses triggered by an acute myocardial infarction (MI) and their possible role in the progress of this disease. Serum samples were collected from two groups of patients, group A (n = 26) within the first week of MI, and group B (n = 28) at 2 weeks and 2 months after MI. Patients in group A were older and had higher prevalence of hypertension and previous attack of MI than patients in group B. The levels of anti-myosin immunoglobulin M and immunoglobulin G antibodies in the serum samples from group A were significantly higher than those in normal control subjects. In group B, the levels of both antibodies were lower than those in group A but remained significantly higher than those in normal control subjects at both 2 weeks and 2 months. The levels of intercellular adhesion molecule-1 (sICAM-1) and vascular cell adhesion molecule-1 (sVCAM-1) in the serum samples from group A patients were significantly higher than those in normal control subjects. At 2 weeks after MI (group B), only the level of sVCAM-1, but not that of sICAM-1, was significantly higher than that in normal control subjects, and there were no significant changes in the levels of these two molecules from 2 weeks to 2 months after MI. We conclude that the higher levels of anti-myosin antibodies and adhesion molecules in group A patients as compared with group B patients may be due to higher or more frequent exposures of their immune systems to heart antigens. Furthermore, the immunoglobulin M antibody response during the first week of MI had an inverse relationship with the level of interleukin-2R (sIL-2R), which suggested a possible suppressive or regulatory role of this antibody on the cellular immune response during this time.

Affine-similar shape retrieval: application to multiview 3-D object recognition.

The feasibility of representing a three-dimensional (3-D) object with a small number of standard views is studied. The object boundary of each view is considered as a two dimensional (2-D) shape and is represented by the locations of the maxima of its curvature scale space (CSS) image contours. The idea is to identify an unknown object from an image taken from a random view by using the stored descriptions of the standard views. The CSS image has been selected for MPEG-7 standardization. The maxima of CSS image have already been used to represent 2-D shapes in different applications under similarity transforms. Since the new application involves affine transforms, we first examine the effects of general affine transforms on the representation and show that the locations of the maxima of the CSS image do not move dramatically even under large affine transformations. Our system for shape-based retrieval from large image databases is then applied to multiview 3-D object representation and recognition. Our collection of 3-D objects consists of 18 aircraft of different shapes. Three silhouette contours corresponding to random views are separately used as input for each object. Results indicate that robust and efficient 3-D free-form object recognition through multiview representation can be achieved using the CSS representation.

B cells and antibodies in the pathogenesis of myelin injury in Semliki Forest Virus encephalomyelitis.

To determine the contribution of B cells to brain myelin injury in Semliki Forest Virus (SFV) encephalomyelitis, normal C57BL/6 (B6) and B-cell-deficient (C57BL/6-tm1Cgn) B6 mice were infected with SFV. The peak of clinical disease, i.e., the time at which the greatest proportions of mice had moderate to severe clinical signs, appeared earlier in B6 mice [day 7 postinfection (pi)] than in B-cell-deficient mice (day 21 pi). By flow cytometry, no clear differences were found in the percentages of CD3(+)CD4(+) T cells in the brains of B6 and B-cell-deficient mice. However, by day 21 pi, percentages of CD3(+)CD8(+) T cells were greater in brains of B-cell-deficient than in those of B6 mice. On day 21 pi, percentages of CD19(+) B cells were maximal in B6 mice, but B cells were absent in B-cell-deficient mice at all time points. Sera obtained from B6 mice showed antibody responses to SFV, to SFV E2 peptides p137-151 and p115-133, and to peptides of myelin oligodendrocyte glycoprotein p18-32 and myelin basic protein (MBP) p64-75. Sera obtained from B-cell-deficient mice showed minimal or no reactivity to SFV, E2, or myelin peptides. CNS inflammatory and PAS-positive macrophage foci were maximal on days 7-14 pi in all mice. Additionally, B6 mice had brain white matter vacuolation, whereas B-cell-deficient mice did not. These data suggest that brain infiltrating B cells and anti-myelin antibodies contribute to myelin injury in SFV encephalomyelitis.

Lymphocyte activation in angina pectoris.

To determine the role of immune responses in the etiology of coronary angioplasty, the distribution of blood lymphocytes and levels of soluble immune factors in sera of patients with primary unstable angina were determined at pre and post coronary angioplasty. Our data showed (1) an increase in the numbers of lymphocytes bearing lymphocyte activating gene-3 (LAG-3) and CD40 in the blood and (2) an increase in levels of sIL2-R and sVCAM-1 in the sera of patients with unstable angina, compared with normal subjects. In contrast, there were no changes in these values in blood or sera of patients shortly after coronary angioplasty. However, levels of sCD8 in the sera of patients, which were similar to those of normal subjects, significantly increased post coronary angioplasty. These results indicate that peripheral blood lymphocytes of patients with unstable angina are immunologically activated and are producing soluble factors which may allow their interaction with endothelial cells in areas of inflammation. This may play a role in antigen presentation and T-B cell interactions which can lead to potentiation of heart disease.

Molecular mimicry between a viral peptide and a myelin oligodendrocyte glycoprotein peptide induces autoimmune demyelinating disease in mice.

Semliki Forest Virus (SFV) induces an encephalomyelitis followed by demyelination in the brains of C57Bl6/J (B6) mice. To investigate the role of molecular mimicry in the pathogenesis of postviral demyelination, alignment algorithms were used and amino acid homologies between immunogenic epitopes of SFV and myelin autoantigens, myelin basic protein (MBP), myelin proteolipid protein (PLP) and myelin oligodendrocyte glycoprotein (MOG) were identified. Immunization of B6 mice with SFV proteins induced significant lymphocyte proliferation to SFV E2 peptides and to MOG peptide, 18-32 (which had molecular mimicry with E2 115-129), but not to MBP or PLP peptides. Both MOG 18-32 and E2 115-129, induced a later-onset chronic EAE-like disease that correlated with the presence of multifocal vacuolation in the CNS white matter. This histopathology was reminiscent of the secondary demyelination seen following SFV infection. Serum antibody responses to the peptides appeared late after immunizations and some samples cross-reacted with other myelin peptides, as well as with the mimicked MOG peptides. These findings suggest that following a CNS viral infection, antibody response to an epitope of virus that exhibits molecular mimicry with a peptide of MOG may contribute to autoimmune mediated injury to CNS myelin.

Influenza virus vaccination of patients with multiple sclerosis.

Prior to vaccination with a trivalent influenza vaccine (AT/Texas, AB/Beijing, and BP/Panama), sera from 19 MS patients had a significantly higher mean level of antibody than 9 normal subjects to AT strain of influenza, but not to AB or BP strains. After Flu vaccination, the mean anti-AT and anti-AB antibody titers significantly increased 4-fold in 11 MS patients and 9 normal subjects. The ratio of MS responders (6/11), however, was lower than normal (8/9). The mean PBL proliferative response to the Flu antigens increased after vaccination significantly more in MS patients than in normal subjects, and increased in 9 of 11 MS patients and 3 of 9 normal subjects. Although MS patients responded to Flu antigens with higher antibody levels and proliferative responses of PBL, than normal subjects, a clinical protective effect of the vaccine against Flu was not clearly demonstrated in these patients, and vaccination did not cause or protect against exacerbation of MS.

Production and role of cytokines in the CNS of mice with acute viral encephalomyelitis.

Semliki Forest Virus (SFV) causes a more severe acute encephalomyelitis in B6 than in SJL mice despite similar T cell proliferation and antibody responses in these two strains. To determine the immunological mechanisms that may contribute to this difference, CNS tissues from SFV-infected B6 and SJL mice were analyzed for viral replication, inflammatory responses and cytokine production, by semiquantitative reverse transcriptase-PCR and immunohistochemistry. Although initially similar on day 2 p.i., SFV replicated to higher viral titers in B6 than SJL mice on days 4 and 7 p.i. Infectious virus was cleared from both strains by day 10 p.i. There were no differences in numbers of CD4+, CD8+ or MHC class I and II+ inflammatory cells at any time point. Higher levels of IL-4 mRNA, lower levels of TNF-alpha, IL-6, IL-1 beta and IL-2 mRNAs and lower IL-2+ and IFN-gamma+ cells were found in B6. These findings suggest that despite comparable immune responses, different patterns of cytokine production correlated with higher levels of virus in the brains and more severe clinical disease in B6, and more efficient clearance of virus and less severe disease in SJL mice.

Treatment of myasthenia gravis by immunoadsorption of plasma.

We treated 16 patients with moderately severe to severe generalized myasthenia gravis (MG) by immunoadsorption (perfusion through a resin that adsorbs proteins) of 2,500 ml plasma on each of four alternate days. Fourteen patients who completed treatment all had significant improvement in strength (6 excellent, 6 good, and 2 fair), which began a mean of 42 hours after the first immunoadsorption, reached a maximum 4 days after the fourth immunoadsorption (mean, 250% of baseline strength), and returned to baseline over a mean of 2 months. Thirty-seven grams of plasma proteins were removed during each immunoadsorption, which required no replacement, compared with 175 grams during plasma exchange, which requires replacement with albumin. Serum or plasma concentration of all proteins fell, more so for most of the larger proteins than for the smaller ones: acetylcholine receptor antibody (AChR Ab) fell to a mean of 23% of original level, fibrinogen to 26%, C4 to 29%, IgM to 33%, IgG to 35%, CH50 to 41%, C3 to 42%, IgA to 54%, and albumin to 76%. All proteins, including AChR Ab, returned to their original levels within 1 to 3 weeks after the last immunoadsorption, while improvement in strength lasted a mean of 6 weeks longer. One seronegative patient had excellent improvement lasting more than a month. Activated complement C5a and white blood cell count rose during each immunoadsorption, while activated complement C3a fell, and each returned to its original level within hours. Eight patients had transient symptomatic hypotension attributable to withdrawal of blood more rapidly than it was returned; this hypotension was prevented or ameliorated by intravenous saline.(ABSTRACT TRUNCATED AT 250 WORDS)

Immune responses, and autoimmune outcome, during virus infection of the central nervous system.

A combined role of a virus infection of the central nervous system (CNS) and an autoimmune response to myelin basic protein (MBP), an autoantigen of the CNS, is suggested in the pathogenesis of multiple sclerosis (MS). SJL mice are highly susceptible while B6 mice are less susceptible to the induction of experimental autoimmune encephalomyelitis (EAE), the autoimmune model of MS. Peripheral inoculation of Semliki forest virus (SFV) into SJL and B6 mice resulted in: (1) Higher viral titers, more severe clinical disease, and hence a stronger nonspecific and SFV-specific lymphoproliferation, and production of IFN-gamma and TNF/LT was observed by splenocytes (SPL) of B6 than by those of SJL mice, on Day 7 postinfection. (2) Following viral clearance, however, proliferation to SFV, and to MBP, and the production of IFN-gamma and TNF/LT by SPL of SFV-infected SJL mice were significantly higher, while the production of TGF-beta was significantly lower than by those of B6 mice. In conclusion, the immune responses to SFV, and to MBP, which were triggered by SFV infection were significantly higher and more prolonged in the SPL of SJL mice, the EAE-susceptible mice, than by those of B6 mice after the infection was cleared.

Production of anti-acetylcholine receptor-alpha antibody in vitro by peripheral blood lymphocytes of patients with myasthenia gravis: role of immunoregulatory T cells and monocytes.

To study the role of T cells in T and B cell interaction resulting in production of antibody (Ab) to the alpha chain of acetylcholine receptor (anti-AChR-alpha Ab) in myasthenia gravis (MG), we cocultured peripheral blood-purified B and T cells of patients with MG and of control subjects with and without multiple sclerosis in the presence of AChR-alpha or pokeweed mitogen. Under these conditions, a high level of anti-AChR-alpha Ab was produced by cells of patients with MG but not of control subjects. Production of anti-AChR-alpha Ab by B cells was stimulated by autologous purified or cloned CD4+ T cells, whereas autologous CD8+ T cells had no effect. CD8+ T cells did not suppress anti-AChR-alpha Ab production when added to B cells cocultured with CD4+ T cell clones. Anti-AChR-alpha Ab production was inhibited by monoclonal antibodies against CD4 and class II major histocompatibility complex (MHC) antigens, indicating that these antigens are required for productive T-B cell interactions resulting in anti-AChR-alpha Ab synthesis. Anti-AChR-alpha Ab production by peripheral blood lymphocytes of patients with MG was significantly lower than that by their purified or cloned T cells cultured with B cells. Cell-mixing experiments indicated that anti-AChR-alpha Ab synthesis was inhibited by monocytes. The prostaglandin synthetase inhibitor, indomethacin, partially restored the suppressive effect of monocytes on anti-AChR-alpha Ab synthesis. These results indicate that induction of anti-AChR-alpha Ab production by CD4+ T cell clones requires CD4 and class II MHC antigens and is inhibited by suppressor macrophages and not by CD8+ T cells.

Defective production of anti-inflammatory cytokine, TGF-beta by T cell lines of patients with active multiple sclerosis.

Activated T lymphocytes play an important role in the pathogenesis of multiple sclerosis (MS). These T cells secrete both pro- and anti-inflammatory cytokines. We have studied the production of these two kinds of cytokines by PBL of patients with MS and compared it with normal controls and other autoimmune diseases (OAD). PBL of 29 patients with MS, 14 patients with OAD, and 14 healthy normal controls were cultured for 5 wk. PBL of MS patients produced more pro-inflammatory cytokines, IL-2, IFN-gamma and TNF/lymphotoxin, and less anti-inflammatory cytokine, TGF-beta, during wk 2 to 4 in culture than PBL of normal controls. PBL of MS patients also produced more IL-2 and TNF/lymphotoxin than PBL of OAD patients. Decreased TGF-beta production by lymphocytes of patients with MS correlated directly with disease activity. MS patients with active disease produced less TGF-beta than MS patients with stable disease. The cells producing TGF-beta were primarily CD8+ T cells and CD45RA+T cells. These findings emphasize the complexity of immune response in MS patients and suggest that the increased production of pro-inflammatory cytokines by lymphocytes of patients with MS, combined with the decreased production of TGF-beta (anti-inflammatory cytokine), may play an important role in the mechanisms and manifestations of MS.

Presence of cross-reactive antibodies to HTLV-1 and absence of antigens in patients with multiple sclerosis.

Antibodies to HTLV-1, as determined by ELISA, were highly elevated in the serum samples of four out of four (100%) patients with TSP, moderately elevated in four out of four (100%) HTLV-1 carriers, slightly elevated in 12 out of 34 (35%) patients with MS, and absent from the serum samples of 34 normal subjects. Western blot analysis showed that the antibodies to HTLV-1 antigens in MS serum were heterogeneous. Cultivation of peripheral blood lymphocytes (PBLs) from patients with MS or normal subjects did not generate HTLV-1 core p19 antigen in the supernatant of culture medium, whereas cultivation of PBLs from patients with TSP and carriers of HTLV-1 generated core p19 antigen after 3 days for up to 28 days of cultivation. HTLV-1 antigens were also expressed on the surface of PBLs in three out of four patients with TSP and in two out of four HTLV-1 carriers on days 14 and 28 of cultivation, as measured by indirect immunofluorescence or alkaline phosphatase staining, but were not found in PBLs of any of 34 patients with MS or 34 normal subjects. The data indicate that although cross-reacting antibodies appear in the serum of some patients with MS, not enough evidence exists to suggest that HTLV-1 antigen is being produced in MS or that HTLV-1 plays a role in the pathogenesis of this disease.

Needlestick injury: blood, mononuclear cells, and acquired immunodeficiency syndrome.

INTRODUCTION: Because transmission of HIV to health care workers after needlestick injury has occurred mainly a result of deep insertion of large gauge needles, blood and viable mononuclear cells transferred after needlestick injury were measured. METHODS: Needles of 20 to 27 gauge were filled with HIV-1 seropositive blood and inserted through extracorporeal human skin or parafilm covering physiologic saline solution modified Drabkin's solution, or culture medium, or inserted directly into one of these fluids, to a depth of one third of the needle length (0.5 inch) for 1 second. Volume of blood transferred was measured by both modified Drabkin's method and by chromium 51 labeling of red blood cells. Transfer of viable mononuclear cells was measured by growth in culture medium containing autologous feeder cells. RESULTS: The volume of blood transferred from a needle passed through skin varied from 312 +/- 69 nl from a 20-gauge needle to 14 +/- 4 nl from a 27-gauge needle, as measured by modified Drabkin's technique, and from 404 +/- 80 nl to 12 +/- 3.1 nl, as measured by chromium 51 labeling of red blood cells. The volume of blood transferred from a needle passed through parafilm was twice that transferred through skin. The volume of blood transferred through skin was 40% that transferred directly into fluid not covered by any barrier; blood transferred through parafilm was 80% of that transferred directly. When needles containing blood were inserted into culture medium for 1 second in the absence of a barrier, at least one viable mononuclear cell was almost always transferred to fluid from all gauges of needle tested. Insertion of needles through skin prevented transfer of all viable mononuclear cells from only 3% to 5% of 20- to 23-gauge needles, and from 12% to 32% of 26- and 27-gauge needles. Parafilm was an even less effective barrier than skin. Insertion of needles through parafilm completely prevented transfer of viable mononuclear cells from no 20- to 23-gauge needles and from only 5% to 10% of 26- and 27-gauge needles. CONCLUSION: The volume of blood transferred after needle insertion through skin for 1 second varied with the gauge of the needle and was 30-fold higher from a 20-gauge than from a 27-gauge needle. Variable mononuclear cells were transmitted after insertion through skin from more than 95% of 20- to 23-gauge needles and from 68% to 88% of 26- and 27-gauge needles. Parafilm was less effective than skin in reducing transmission of blood and viable mononuclear cells.

Characterization of a T suppressor cell line that downgrades experimental allergic encephalomyelitis in mice.

A T-suppressor (Ts) cell line of CD8 phenotype was isolated from spleens of SJL/J mice that had recovered from experimental allergic encephalomyelitis (EAE) induced by injection of MBP-activated T cells. The Ts cell line inhibited the proliferation of MBP-sensitized T cells in vitro. Addition of recombinant IL-2 enhanced the Ts-mediated suppression. Adoptively transferred Ts line was able to downgrade EAE in mice subsequently challenged with MBP-activated T cells. The mechanism of suppression appeared to involve neither direct cytolysis of the effector T cells nor the production of a soluble suppressor factor. The findings suggest an in vivo role for suppressor T cells in the regulation of EAE.

Characterization of in vivo-activated T cell clones from peripheral blood of multiple sclerosis patients.

In vivo-activated interleukin-2 responsive T cell clones were generated from peripheral blood (PBL) of multiple sclerosis patients (MS) and normal subjects (N) by limiting dilution analysis. The frequency with which interleukin-2 responsive cells were cloned from PBL was higher in MS than N. CD8 was the predominant phenotype expressed by both MS (85%) and N (89%) clones. Seven clones from four MS patients but none from five N subjects specifically proliferated against myelin basic protein. These studies demonstrate the existence of MBP-reactive T cells in PBL of MS patients.

Phenotypic and functional characterization of T cells from patients with myasthenia gravis.

A study of cell surface phenotypes of PBL of myasthenia gravis (MG) patients showed that their T cells had a significantly higher percentage of 4B4+ T cells (the helper/inducer subset) than age- and sex-matched controls. The PBL of MG patients proliferated significantly higher than those of normal subjects (NS) in response to the purified alpha chain of the acetylcholine receptor (AChR). Anti-AChR antibody was present in sera of 88% of MG and none of the NS. The PBL B cells from MG only, when cultured with autologous T cells and stimulated with either pokeweed mitogen (69%), or AChR-alpha chain (38%), secreted antibody to AChR-alpha chain, whereas T and B cells alone secreted no antibody. T cells from PBL of MG patients were more readily cloned than T cells of NS, by limiting dilution, in the presence of recombinant IL-2 and in the absence of AChR-alpha chain. About 50% of T cell clones from MG patients, compared to none from NS, proliferated to AChR-alpha chain. This response was HLA-DR restricted. MG T cell clones did not display significant cytotoxic activity, as compared to control T cell clones. Our results indicate that in MG, 4B4+ regulatory T cells play their role in the pathogenesis of MG, not by cytotoxicity, but more likely by their ability to stimulate specific antibody production by B cells.

Detection of HIV antibody and antigen (p24) in residual blood on needles and glass.

There is a significant rate of percutaneous injury with needles during the care of patients with acquired immunodeficiency syndrome (AIDS). Following puncture injury, it is recommended that the source of the contaminating blood be checked, and if human immunodeficiency virus-type 1- (HIV-1)-seropositive, zidovudine prophylaxis be considered. As the source of contaminating blood may be unknown, we studied the detectability of HIV-1 antibody and circulating antigen (p24) in the residual blood from needles and pieces of glass at various intervals following exposure to blood. The residual volume of blood remaining in needles varied from 183 +/- 50 microliters for a 20 G needle to 7.8 +/- 1 microliter for a 27 G needle, and the residual blood on small pieces of glass varied from 23 microliters for a piece weighing 558 mg to 2 microliters for a piece weighing 21 mg. Analysis of washed samples of residual blood from all 20 G through 26 G needles and from broken pieces of glass larger than 0.41 g that had been exposed to HIV-1-seropositive blood and left at room temperature for one hour, one day and one week resulted in positive tests for HIV-1 antibody by enzyme-linked immunosorbent assay (ELISA), immunofluorescence and Western blot assays. The circulating antigen was detected in residual blood of 20 G through 26 G needles, but not from contaminated pieces of glass. This technique could be applied to situations where a healthcare worker pricked him- or herself with a needle or with a piece of glass that had been contaminated with blood of unknown seroreactivity.(ABSTRACT TRUNCATED AT 250 WORDS)

Role of the immune response in Sindbis virus-induced paralysis of SJL/J mice.

The pathologic role of the specific immune and inflammatory responses to viral infections of the CNS was investigated by using mice which are susceptible (SJL/J) and resistant (C57Bl6 and BALB/c) to the development of experimental autoimmune encephalomyelitis (EAE). Intracerebral inoculation of 10(4) PFU of Sindbis virus (SV) into 6- to 8-wk-old SJL/J mice resulted in a severe and sometimes fatal encephalomyelitis. A mild to severe hind leg paralysis was observed around days 6 to 7 postinfection (pi) which closely resembled EAE stages and persisted for up to 8 wk pi. Immunosuppression with cyclophosphamide on day 4 alleviated the severity of this disease. Significant perivascular and parenchymal infiltration was present in the brains and spinal cords of SV-infected SJL/J mice for up to 1 mo. This apparent immunopathologic reaction was found to be a characteristic of SJL/J mice, because infection of 6- to 8-wk-old BALB/c and C57Bl6 mice with SV did not cause paralytic disease. These mice also exhibited a significantly milder cellular infiltrate which was mostly resolved on day 12 to 14 pi. Titers of virus in the brain and spinal cords of mice were comparable with clearance by day 7 pi. SV-specific lymphoproliferation and serum antibody responses were also comparable in all mice. SV-infected SJL/J mice developed antibodies to myelin components as demonstrated in Western blots and responded to myelin basic protein by lymphoproliferation. Lymph node cells from these mice, after in vitro challenge with myelin basic protein, transferred a mild EAE-like disease to naive recipients and potentiated subclinical EAE into a severe disease.