Oligodendrocyte-specific expression and autoantigenicity of transaldolase in multiple sclerosis.

Although the etiology of multiple sclerosis (MS) is unknown, there is compelling evidence that its pathogenesis is mediated through the immune system. Molecular mimicry, i.e., crossreactivity between self-antigens and viral proteins, has been implicated in the initiation of autoimmunity and MS. Based on homology to human T cell lymphotropic virus type I (HTLV-I) a novel human retrotransposon was cloned and found to constitute an integral part of the coding sequence of the human transaldolase gene (TAL-H). TAL-H is a key enzyme of the nonoxidative pentose phosphate pathway (PPP) providing ribose-5-phosphate for nucleic acid synthesis and NADPH for lipid biosynthesis. Another fundamental function of the PPP is to maintain glutathione at a reduced state and, consequently, to protect sulfhydryl groups and cellular integrity from oxygen radicals. Immunohistochemical analyses of human brain sections and primary murine brain cell cultures demonstrated that TAL is expressed selectively in oligodendrocytes at high levels, possibly linked to production of large amounts of lipids as a major component of myelin, and to the protection of the vast network of myelin sheaths from oxygen radicals. High-affinity autoantibodies to recombinant TAL-H were detected in serum (25/87) and cerebrospinal fluid (15/20) of patients with MS. By contrast, TAL-H antibodies were absent in 145 normal individuals and patients with other autoimmune and neurological diseases. In addition, recombinant TAL-H stimulated proliferation and caused aggregate formation of peripheral blood lymphocytes from patients with MS. Remarkable amino acid sequence homologies were noted between TAL-H and core proteins of human retroviruses. Presence of crossreactive antigenic epitopes between recombinant TAL-H and HTLV-I/human immunodeficiency virus type 1 (HIV-1) gas proteins was demonstrated by Western blot analysis. The results suggest that molecular mimicry between viral core proteins and TAL-H may play a role in breaking immunological tolerance and leading to a selective destruction of oligodendrocytes in MS.

Comparative analysis of antibody and cell-mediated autoimmunity to transaldolase and myelin basic protein in patients with multiple sclerosis.

Antibody and T cell-mediated immune responses to oligodendroglial autoantigens transaldolase (TAL) and myelin basic protein (MBP) were examined in patients with multiple sclerosis (MS). Immunohistochemical studies of postmortem brain sections revealed decreased staining by MBP- and TAL-specific antibodies in MS plaques, indicating a concurrent loss of these antigens from demyelination sites. By Western blot high titer antibodies to human recombinant TAL were found in 29/94 sera and 16/23 cerebrospinal fluid samples from MS patients. Antibodies to MBP were undetectable in sera or cerebrospinal fluid of these MS patients. Proliferative responses to human recombinant TAL (stimulation index [SI] = 2.47+/-0.3) were significantly increased in comparison to MBP in 25 patients with MS (SI = 1.37+/-0.1; P < 0.01). After a 7-d stimulation of PBL, utilization of any of 24 different T cell receptor Vbeta gene segments in response to MBP was increased less than twofold in the two control donors and six MS patients investigated. In response to TAL-H, while skewing of individual Vbeta genes was also less than twofold in healthy controls, usage of specific Vbeta gene segments was differentially increased ranging from 2.5 to 65.9-fold in patients with MS. The results suggest that TAL may be a more potent immunogen than MBP in MS.

The prognostic significance of neuroendocrine markers and carcinoembryonic antigen in patients with resected stage I and II non-small cell lung cancer.

Non-small cell lung cancer with neuroendocrine differentiation may represent a subset of patients with a more aggressive (like small cell lung cancer) or less aggressive (like carcinoid) biological behavior. To investigate their prognostic significance, immunohistochemical stains for 4 neuroendocrine markers (neuron-specific enolase, chromogranin A, Leu-7, and synaptophysin) and carcinoembryonic antigen (CEA) were studied in 260 patients with surgically resected stage I and II non-small cell lung cancer. The following percentages of cases were positive for each marker: neuron-specific enolase, 70.0%; chromogranin A, 14.2%; Leu-7, 7.7%; synaptophysin, 11.2%; and CEA, 68.5%. Sixty-one (23.5%) were positive for > or = 2 neuroendocrine markers. When compared to adenocarcinoma, squamous cell carcinoma displayed lower positivity for CEA and > or = 2 neuroendocrine markers. There was no significant difference in stage, site of relapse (distant versus local), disease-free, or overall survival for each marker individually or for those with > or = 2 neuroendocrine markers. Multivariate analysis showed that higher nodal stage (N1 versus N0), tumor stage (T2 versus T1), older age, and the presence of mucin predicted for poorer overall survival. Neuroendocrine markers and CEA were not of prognostic significance in this group of patients with resected stage I and II non-small cell lung cancer.

Blood group antigen A and flow cytometric analysis in resected early-stage non-small cell lung cancer.

The loss of blood group antigen A on tumor tissue has been reported to be a strong adverse prognostic marker for patients with resected non-small cell lung cancer (NSCLC). Results have varied with respect to the prognostic significance of flow cytometric data. We sought to confirm the prognostic significance of blood group antigen A loss and flow cytometry in a large cohort of patients with early-stage NSCLC. Two hundred and sixty patients with surgically resected stage I (n = 193) and II (n = 67) NSCLC with at least a 5-year follow-up were identified. Using paraffin-embedded primary tumor, immunohistochemical stains for blood group antigen A were performed on 90 patients with blood type A or AB. The DNA index and percentage of cells in S phase were successfully obtained on 188 and 152 patients, respectively. The median survival time of the patients with primary tumors negative for blood group antigen A was 38 months (n = 36), compared with 98 months (n = 54) for those with antigen A-positive tumors (P < 0.01). The median disease-free survival times for antigen A-negative and -positive tumors were 26 months and 98 months, respectively (P < h 0.01). The median survival time of the patients with aneuploid tumors was 51 months (n = 131), compared with 50 months (n = 57) for those with diploid tumors (P = 0.42). The median survival time of the patients with S phase >8% was 44 months (n = 105), compared with 60 months (n = 47) for those with S phase

erbB-2 and myc oncoproteins in sera and tumors of breast cancer patients.

This study compares the prevalence of elevated serological levels of erbB-2 and myc proteins in 36 breast cancer patients and 25 healthy, ambulatory female controls. The controls were frequency matched to the cases by age and ethnicity. Oncoprotein levels were determined blind to the "case-control status" of the individual from whom the specimen was derived. Corresponding tissue levels were examined in tumors of the 13 cases from whom sufficient tissue was available. Serum oncoproteins were elevated as follows: erbB-2 in one control (4%) compared with nine cases (25%; PFisher's exact = 0.03); myc in no control (0%) compared with seven cases (19%; PFisher's exact = 0.02). Elevated serum levels of erbB-2 or myc oncoproteins were detected in four of the seven cases (57.1%) of in situ cancer without evidence of infiltration. In all cases with elevated serum oncoproteins where tumor tissue was available, the corresponding protein was elevated in the tumor. The three cases who had elevated preoperative serum oncoprotein levels and from whom it was possible to procure postoperative specimens had normal postoperative serum oncoprotein levels. We conclude that (a) erbB-2 and myc oncoproteins are elevated in a proportion of breast cancer patients, (b) the tumor seems to be the source of the serum elevation, and (c) these proteins may be useful as part of a panel of biomarkers of early malignant disease.

Lyme disease patients' serum contains IgM antibodies to Borrelia burgdorferi that cross-react with neuronal antigens.

Serum from patients with neurologic manifestations of Lyme disease had serum IgM antibodies that bound to normal human axons, whereas binding was absent or weak in patients without neurologic findings. Antiaxonal binding could be eliminated by absorption with Borrelia burgdorferi. A murine monoclonal antibody to the borrelial flagellin also bound to human axons.

Large scale recovery of biologically active IgM (95% pure) from human plasma obtained by therapeutic plasmapheresis.

We have developed a large scale (10 g/batch) non-chromatographic method for the collection of IgM directly from plasma obtained from therapeutic plasmapheresis of patients. The technique uses sequential precipitations from ammonium sulfate and polyethylene glycol, and gives high yields of undenatured IgM antibodies. The biological activity of IgM prepared by this method has been demonstrated by systemically transfusing animals with the antibody and producing a demyelinating syndrome very similar to the antibody donor. Control IgM preparations exhibited no toxicity after daily injections of antibody for 8 weeks, showing that the method gives antibody with excellent bio-compatibility (Tatum, 1989). The method, which is effective for monoclonal and polyclonal antibodies, is useful when large amounts of antibodies are needed for passive transfer or other studies.

Rapid serologic diagnosis of serum sickness from antithymocyte globulin therapy using enzyme immunoassay.

Although the use of antithymocyte globulin/antilymphocyte serum (ATG/ALS) has been shown to be beneficial in treating renal allograft rejection, the incidence and nature of serum sickness reactions following such treatment have received limited attention. In the setting of rejection or infection, the diagnosis of serum sickness is often difficult on clinical grounds, and biopsy may be an undesirable means of obtaining documentation. A sensitive enzyme-linked immunosorbent assay (ELISA) was developed to detect antibodies against ATGAM (Upjohn) and was utilized in 35 patients treated for rejection with ATGAM following clinical unresponsiveness to bolus methylprednisolone therapy. Serum sickness was diagnosed clinically in 7 of these patients (20%) during or immediately following ATGAM therapy. Significant titers of anti-ATGAM antibody were found in these 7 cases, as well as 5 additional cases (14%). Upon retrospective review, the diagnosis of serum sickness was also made in each of these 5 additional cases based upon the immunopathologic detection of horse immunoglobulin deposits in vessels of tissue (lung or kidney) examined. In 2 other cases where serologic testing was negative, there was some clinical suggestion of serum sickness that could not be documented pathologically. ELISA for anti-ATGAM antibodies was negative in testing 46 control patients and pooled normal sera. In one case, retrospective testing of pre-ATGAM therapy serum samples showed significant antihorse antibodies. Despite a negative skin test prior to administration, this patient developed symptoms of serum sickness 15 days after the onset of ATGAM therapy. Extremely high titers of antibody were detected 5 days earlier (10 days posttherapy), and the presence of horse immunoglobulin immune complexes in the kidney was documented following graft loss that occurred within 4 weeks. These findings indicate (1) serologic testing for anti-ATGAM antibodies using a sensitive ELISA method provides a quick, inexpensive, noninvasive means for the presumptive diagnosis of serum sickness in complicated clinical situations, and (2) screening of patient sera prior to administration of ATGAM may be useful in avoiding potential serum sickness reactions.

Relationships among the histologic pattern, intensity, and phenotypes of T cells infiltrating renal allografts.

The evaluation of renal allograft rejection by routine histologic evaluation of transplant biopsy is often a diagnostic problem. Advances in monoclonal antibody and immunohistochemical technology have led to their application in characterizing the phenotype of peripheral blood lymphocytes in transplant recipients, as well as the infiltrating lymphocytes in renal biopsy specimens (both tissue and aspiration cytology samples). Recent reports suggest that the relative number of infiltrating T cells and the T cell phenotypic subset ratio may have some association with graft survival. However, most allograft biopsy studies have not examined the relationship of the intensity and phenotype of T cells to the histologic pattern of infiltration, while fine-needle aspiration cytology (FNAC) and peripheral blood analysis do not allow for this assessment. To examine the association of histologic pattern with the intensity and phenotype of T cell infiltrates, immunoperoxidase labeling of cells using monoclonal antibodies was evaluated in 66 renal allograft biopsies performed because of clinical suspicion of rejection. Our findings indicate that the ratio of T helper-inducer (Leu 3) cells to T suppressor-cytotoxic (Leu 2) cells is significantly lower in the pattern of diffuse renal cortex infiltration (cortical diffuse [CD] pattern) when compared with other histologic patterns (cortical aggregate or perivascular), regardless of the overall intensity of the T cell infiltrate. A significant difference in T cell phenotypes was also found among different histologic patterns as expressed by Leu 3/Leu 2 ratios and by the number of Leu 2 cells. An increase in the overall intensity of T cell infiltrate was not associated with significant changes in T cell phenotype seen in any pattern, but an increase in the intensity of the cortical diffuse T cell infiltrate was associated with a significant decrease in the Leu 3/Leu 2 ratio (P less than 0.04), which was due to an increase in the population of Leu 2 cells (P less than 0.002). Interestingly, increases in the intensity of either cortical aggregate or perivascular T cell infiltrates were associated with significant increases in both Leu 3 and Leu 2 cells, as well as a tendency towards higher Leu 3/Leu 2 ratios. These findings indicate that evaluation of T cell infiltrates by phenotype in renal allografts is dependent upon the pattern as well as the intensity of infiltrate.

Affinity studies of human anti-MAG antibodies in neuropathy.

Human IgM anti-myelin associated glycoprotein (MAG) antibodies from patients with neuropathy bind to oligosaccharide determinants shared by MAG and several other glycoconjugates in peripheral nerve. The apparent affinities of human anti-MAG antibodies were determined by an ELISA system which measures free antibody concentration at equilibrium in solution. Intact MAG, which bears multiple antigenic oligosaccharides, and monovalent oligosaccharides generated by pronase digestion of MAG were used as antigen. The human antibodies bound to intact MAG with dissociation constants of between 2.5 x 10(-10) M and 2.1 x 10(-7) M, and to the monovalent oligosaccharides with up to 100-fold lower affinities. Reduction of the pentameric IgM to its monomeric counterpart reduced its affinity to intact MAG 5-fold, but its avidity for immobilized MAG was reduced 500-fold as determined by ELISA. These studies show that IgM Anti-MAG antibodies exhibit relatively low intrinsic affinities for the oligosaccharide antigen, but their affinities and avidities are significantly increased by the multivalent nature of the antibody-antigen interaction.

Adenovirus infection of the renal allograft with sparing of pancreas graft function in the recipient of a combined kidney-pancreas transplant.

We report a case of adenovirus infection of the renal allograft in a combined kidney/pancreas transplant recipient. The clinical presentation was renal allograft failure, which eventually reversed. The pancreatic graft function remained stable. A renal biopsy showed massive tubular necrosis associated with a prominent granulomatous reaction. The process had a striking regional distribution within the kidney with the injury and inflammation limited to the outer medulla. Adenovirus type 11 was isolated from renal tissue by culture, and adenovirus was demonstrated by immunofluorescence and electron microscopy in the kidney biopsy. Immunosuppression may result in unusual patterns of response to infectious agents. This case demonstrated tropism of the adenovirus to only selected tubules within the kidney, with sparing of other organ function including, specifically, the pancreas allograft. The differential diagnosis of a granulomatous reaction in the transplant kidney must be expanded to include viral infection, in particular, adenovirus.

Pathobiology of acute myocardial ischemia: metabolic, functional and ultrastructural studies.

Acute myocardial ischemia induced by coronary occlusion in dogs is most severe in the subendocardial region, whereas more collateral blood flow is often present in the subepicardial region. Initially, all ischemic myocytes are reversibly injured, but beginning at 15 to 20 minutes after the onset, and continuing for 3 to 6 hours, there is a wave front of cell death from the subendocardial region to the less ischemic subepicardial region, such that by 6 hours, the final transmural extent of the infarct is established. Thus, ischemic myocardium cannot be salvaged by reperfusion after greater than or equal to 6 hours of coronary occlusion in open-chest anesthetized dogs. In the severely ischemic subendocardial region, most of the creatine phosphate is lost within the first 3 minutes of ischemia in vivo, and adenosine triphosphate (ATP) is depleted to 35% of control by 15 minutes (when cellular injury is still reversible), and to less than 10% of control at 40 minutes (when injury is irreversible). Tissue ATP content and other indexes of subcellular damage have also been compared after different periods of ischemia using a model of total myocardial ischemia in vitro. As long as the ATP of the tissue was not depleted below 5 mumols/g dry weight, incubated slices of injured myocardium resynthesized high-energy phosphates and excluded inulin. However, lower tissue ATP was associated with depressed high-energy phosphate resynthesis and failure of cell volume regulation. Overt membrane damage, as measured by an increased inulin-diffusible space, was detected only after the tissue ATP decreased to less than 2.0 mumols/g of dry weight. Thus, marked ATP depletion is associated with the onset of structural and functional indexes of irreversible injury. However, whether irreversibility is caused by the marked ATP depletion or by other concomitant metabolic consequences of ischemia is not known. Myocardial ischemic cellular injury is reversible despite depletion of 70% of the control ATP. Nevertheless, when myocyte injury is reversible, there is slow repletion of adenine nucleotides. This slow metabolic recovery may explain the delayed recovery of contractile function observed after reperfusion of ischemic myocardium.

Prognostic significance of Lewis y antigen in resected stage I and II non-small cell lung cancer.

BACKGROUND: The role of Lewis y (Le(y)) antigen expression has been studied extensively in predicting the outcome of various malignancies. We evaluated the expression of Le(y) and its relationship to survival, disease-free survival and other clinicopathologic variables in patients with stage I and II non-small cell lung cancer (NSCLC). OBJECTIVE: To investigate the prognostic significance of Le(y) antigen expression in a large group of well characterized patients with resected stage I and II NSCLC. PATIENTS: Two hundred and sixty patients with surgically resected stage I (n = 193) and II (n = 67) NSCLC with at least 5-year follow-up were identified. RESULTS: The median survival for patients with negative expression of Le(y) (< 50% of cells that were positive) was 46 months, whereas for those with positive expression of Le(y) (> or = 50%), the median survival was 54 months (p = 0.99). The disease-free survival for patients with Le(y)(-) expression was 39 months and 34 months for patients with Le(y)(+) expression (p = 0.3). CONCLUSIONS: We found no relationship between loss of blood group antigen A and expression of Le(y). No statistically significant difference was found in survival between positive and negative expression of Le(y) antigen in patients with resected stage I and II NSCLC.

Pleomorphic fibrohistiocytoma of the breast: a potential pitfall in breast biopsy interpretation.

We describe a benign mammary mesenchymal tumour with atypical stromal giant cells in the contralateral breast of a 66-year-old woman with infiltrating ductal carcinoma. The clinical, morphological and immunohistochemical features of this tumour suggest a pleomorphic variant of fibrous histiocytoma. This benign lesion represents a possible pitfall in breast pathology when interpreting a frozen section or fine needle aspiration biopsy.

Transitional cell carcinoma of renal pelvis: rare occurrence in young male.

Transitional cell carcinoma of the renal pelvis was encountered in a twenty-one-year-old white man. Review of the literature shows this tumor to be rare in individuals less than thirty years of age.

Prognostic markers in resected stage I and II non small-cell lung cancer: an analysis of 260 patients with 5 year follow-up.

We performed a retrospective analysis of potential prognostic markers in 260 patients with surgically resected stage I and II non small-cell lung cancer (NSCLC) with a minimum 5-year follow-up. Cox proportional hazard models and Wilcoxon tests were employed to analyze the effect of patient characteristics on survival and disease-free survival (DFS). In the univariate analysis, the following were significant predictors of shorter overall survival: N-stage (N1 vs N0) (p<0.001); T-stage (T2 vs T1) (p<0.001); antigen A (loss vs presence) (p<0.01); cough (present vs absent) (p=0.01); bcl-2 expression (positive vs negative) (p=0.03); age (>63.5 vs <63.5) (p=0.03); mucin (positive vs negative) (p<0.03). The following were significant predictors of shorter DFS: N-stage (p<0.001); T-stage (p=0.001); loss of antigen A (p=0.01); mucin expression (p<0.01); cough (p=0.02); Ki-67 expression (p=0.02) and negative bcl-2 expression (p=0.03). Analysis of survival difference for histologic subtype, degree of differentiation, aneuploidy, %S-phase, codon 12 K-ras mutation, and immunohistochemistry staining for Lewisy, p53, Rb, microvessel count, HER2, E-cadherin and neuroendocrine markers did not reach statistical significance. In multivariate analysis, the following predicted for shorter overall survival: N-stage (p<0.01), antigen A (p=0.01), age (p<0.01), and bcl-2 (p=0.05); and for DFS, N-stage (p<0.01), antigen A (p<0.01), Ki-67 (p=0.03), mucin (p=0.04) and T-stage (p=0.05). Of all the clinical-pathological, proliferative, and biological markers studied, only a few carried independent prognostic significance.

Scanning electron microscopy and X-ray microanalysis of foreign bodies associated with Silastic implants in humans.

Polydimethylsiloxane (Silastic) capsules containing megestrol acetate have been implanted in subcutaneous tissue as a method of long-term contraception. Histological studies revealed a granulomatous foreign body reaction around these capsules with birefringent crystals in multinucleated giant cells. Although several authors had interpreted these crystals as steroids, this seemed unlikely since the tissue had been processed with organic solvents. Analysis of these crystals by polarization microscopy, scanning electron microscopy, and X-ray microanalysis demonstrated that the material was talc. The talc was probably introduced as a contaminant from gloves during the implantation. Further analysis showed that capsule fragmentation could not have produced the material since Silastic particles could not be detected.

Hypercalcemia with excess serum 1,25 dihydroxyvitamin D in lymphomatoid granulomatosis/angiocentric lymphoma.

Hypercalcemia has been described in a variety of granulomatous and lymphoproliferative disorders in association with elevated serum levels of 1,25-dihydroxyvitamin D. In such cases, hypercalcemia appears to be the consequence of excessive production of 1,25(OH)2D by the lymphocyte/macrophage line. The authors report a patient with lymphomatoid granulomatosis/angiocentric lymphoma who developed hypercalcemia with extreme elevation in serum 1,25(OH)2D. Therapy with steroids reduced the serum calcium and 1,25(OH)2D levels to normal. Hypercalcemia has not previously been reported in lymphomatoid granulomatosis/angiocentric lymphoma. The distinctive features of this malignancy, and the derangement in the metabolism of 1,25(OH)2D in lymphoproliferative disorders in general, are discussed.

HMB-45-positive malignant lymphoma. A case report with literature review of aberrant HMB-45 reactivity.

The monoclonal antibody HMB-45 is a highly specific and sensitive marker of malignant melanoma. Rarely, however, aberrant reactivity to nonmelanocytic tumors has been observed. We report an immunoblastic lymphoma of B-cell phenotype that reacted with HMB-45 in a 70-year-old white woman. To our knowledge, this is the first report of HMB-45-positive malignant lymphoma (excluding plasmacytoma). Of an additional 13 cases of benign and malignant lymphoid tissue, only benign plasma cells in one case reacted with HMB-45.

False-positive antineutrophil cytoplasmic antibody in aspergillosis with oxalosis.

Although highly specific for Wegener's granulomatosis and other vasculitides, false-positive antineutrophil cytoplasmic antibody is occasionally encountered in other nonvasculitic conditions. We report a case of clinically suspected Wegener's granulomatosis with a large pulmonary cavity, renal failure, and positive cytoplasmic antineutrophil cytoplasmic antibody reaction. The patient died before initiation of immunosuppressive therapy. Postmortem examination revealed pulmonary aspergillosis due to Aspergillus niger, with extensive pulmonary and renal oxalosis, which lead to renal failure. The possible role of oxalosis in the pathogenesis of false-positive antineutrophil cytoplasmic antibody reaction is discussed.