Patients with Meniére's disease possess IgE reacting with herpes family viruses.

OBJECTIVE: To determine if patients with Meniere's disease possess serum IgE specific for herpes simplex virus (HSV) type 1, HSV type 2, Epstein-Barr virus, and/or cytomegalovirus. DESIGN: A modified radioallergosorbent test method was employed wherein each serum sample was processed with recombinant protein A to remove competing non-IgE antibodies, and HSV-1, HSV-2, cytomegalovirus, and Epstein-Barr viral proteins were used as potential antigens. PATIENTS: Ten patients with long-standing active Meniere's disease were tested. Ten age- and gender-matched patients with allergic rhinitis but without Meniere's disease served as control subjects. RESULTS: IgE specific for HSV-1, HSV-2, Epstein-Barr virus, and/or cytomegalovirus was found in the serum sample of nine of 10 patients with Meniere's disease but only in four of 10 control serum samples. Of the positive subjects tested, seven patients with Meniere's disease were positive for IgE for at least three viruses compared with only two control subjects. CONCLUSIONS: (1) Most patients with Meniere's disease possess virus-specific IgE in their serum samples; (2) four viruses of the herpes family are capable of inducing such IgE-mediated sensitization; and (3) latent virus-specific, IgE-mediated inflammation may be an important factor in the initiation and/or sustenance of Meniere's disease.

Rapid large-scale growth of Helicobacter pylori in flasks and fermentors.

We developed procedures for large-scale cultivation of Helicobacter pylori in flasks and fermentors. Flasks incubated closed under a microaerophilic gas phase with a cotton plug covered by a plastic bag, followed by removal of the bag after 8 h, gave excellent growth. Growth in a 10-liter fermentor led to excessive foaming if the medium was sparged with gas; silicone- or polyglycol-based antifoaming agents were severely inhibitory. Use of fermentor surface gassing, first with a microaerophilic 6% oxygen gas mixture, then with air, and then with 95% oxygen, allowed the culture to grow to an A600 of 2.5 in < 24 h. This method was modified for scale-up to a 100-liter fermentor.

Serum immunoglobulins specific for intracellular proteins of squamous cell carcinoma.

OBJECTIVE: To determine an autologous humoral immune response to squamous cell carcinoma (SCC) intracellular proteins in patients with SCC. DESIGN: Intracellular proteins were isolated from 25 different cultured SCC lines. The proteins were used as a source of antigens to measure IgA, IgE, and IgG responses in the serum samples of patients and controls. Antibody response was assessed in both unfractionated and fractionated intracellular proteins. PATIENTS: The serum samples of 65 patients with SCC and of 65 age- and gender-matched controls were tested. RESULTS: Antibodies to SCC intracellular proteins were detected in the serum samples of 40 (62%) of the 65 patients with SCC and in the serum samples of 46 (71%) of 65 controls. Thirty (46%) of the patients with SCC and 40 (62%) of the controls had IgE responses, 18 (28%) of the patients and one (2%) of the controls had IgA responses, and 17 (26%) of the patients and 14 (22%) of the controls had IgG responses. An inverse relation was noted between detectable IgE responses and IgA or IgG responses in the patients and the controls. The analysis of antibody response indicated that 28 molecules were recognized predominantly by the serum samples of patients with SCC, but not by the serum samples of controls. CONCLUSIONS: A substantial proportion of patients with SCC and of controls exhibited an autologous humoral immune response to SCC intracellular proteins. The IgE responses to SCC intracellular proteins were inversely related to IgA or to IgG responses. Different antibody isotypes normally cause markedly different immune functions, and may suggest different roles for the existent immune responses to SCC antigens. We identified many tumor-associated antigens that were selectively recognized by the serum samples of patients with SCC. These antigens could be used to define molecular studies of immune surveillance and selection, and may represent appropriate targets for immunotherapy.

Immunity to the HER-2/neu oncogenic protein.

The study of oncogenic viruses led to the discovery that transforming retroviruses contain oncogenes homologous with and/or derived from cellular proto-oncogenes. In humans malignant transformation is often the result of the activation of proto-oncogenes. Normal proto-oncogenes can be activated to transforming proto-oncogenes by a variety of mechanisms including point mutation, translocation and amplification. Development of successful strategies for the immunotherapy of human cancers is an area of intense investigation. Part of the problem in developing cancer-specific immunotherapy has been the lack of well-defined tumour antigens. Our laboratory has focused on the question of whether oncogenic proteins expressed by transforming proto-oncogenes can serve as targets for immune attack. Some patients with HER-2/Neu-positive breast cancer have an existent immune response to the HER-2/neu protein with no clinical signs of autoimmunity, supporting the idea that overexpressed oncogenic proteins can be targeted in therapy without fear of destructive autoimmunity. The identification of candidate cytotoxic T lymphocyte epitopes might allow the generation of tumour-specific cytotoxic T lymphocytes for use in therapy and identify potential epitopes for peptide vaccines.

Existent T-cell and antibody immunity to HER-2/neu protein in patients with breast cancer.

The HER-2/neu protooncogene is amplified and overexpressed in 20-40% of invasive breast cancers. HER-2/neu protein overexpression is associated with aggressive disease and is an independent predictor of poor prognosis in several subsets of patients. The protein may also be related to cancer formation, with overexpression being detectable in 50-60% of ductal carcinomas in situ. It has been suggested that it might be possible to develop specific T-cell therapy directed against proteins involved in malignant transformation. One question is whether normal proteins that are overexpressed are appropriate targets for therapeutic immune attack. This report demonstrates that some patients with HER-2/neu-positive breast cancers have both existent CD4+ helper/inducer T-cell immunity and antibody-mediated immunity to HER-2/neu protein. Initial studies performed on 20 premenopausal breast cancer patients identified antibodies to HER-2/neu in 11 individuals. Similar antibody responses have been found in some normal individuals. The patient with the greatest antibody response was studied in detail. In addition to a humoral immune response this patient had evidence of a significant proliferative T-cell response to the HER-2/neu protein and peptides. Similar T-cell responses have been detected in additional patients. It has been assumed that patients would be immunologically tolerant to HER-2/neu as a self-protein and that immunity might be difficult to generate. If immunity could be generated, the result might be destructive autoimmunity. The current data support the notion that HER-2/neu-specific immunity might be used in therapy without destroying normal tissue but also raises questions as to the role of existent immunity in immune surveillance and cancer progression.

Bacterial allergy in nasal polyposis. A new method for quantifying specific IgE.

OBJECTIVES: To determine (1) if bacteria-specific serum IgE levels can be more effectively measured by first absorbing competing IgG antibodies from serum and (2) if patients with chronic paranasal sinus disease exhibit a high positive prevalence of bacteria-specific serum IgE. DESIGN: A modified radioallergosorbent test method was employed wherein each serum sample was absorbed with recProtein A to remove competing non-IgE antibodies, and purified proteins extracted from 16 individual bacteria were used as potential allergens. PARTICIPANTS: Twenty-four patients with nasal polyposis and 14 with chronic sinusitis, all refractory to conventional medical therapy and requiring endoscopic sinusotomies, were tested. Tested as controls were 10 subjects with chronic allergic rhinitis, without a history of chronic sinus disease, and possessing total serum IgE and inhalant-specific IgE levels equal to or higher than the patient group. RESULTS: (1) Pretreatment of serum samples with recProtein A resulted in an increase of bacteria-specific radioallergosorbent test sensitivity. (2) Seventeen of 24 patients with polyps, eight of 14 with chronic sinusitis, and one of 10 with chronic allergic rhinitis were determined to be IgE positive when tested with this assay. CONCLUSIONS: (1) Bacteria-specific serum IgE can be quantified; (2) most patients with nasal polyposis and/or chronic sinusitis possess bacteria-specific IgE in their serum, while subjects with only allergic rhinitis do not; and (3) multiple bacterial species isolated from chronically infected sinuses are capable of inducing IgE-mediated sensitization.

Plaque-associated immune reactivity as a tool for the diagnosis and treatment of atherosclerosis.

In summary, we have confirmed that a specific humoral immune reactivity involves human atherosclerotic plaque. By isolating some of the autoantigens and employing them in prototype immunoassays, we have been able to measure for the presence of plaque-specific antibodies. Extracted plaque antigens have also served as immunogens in monoclonal antibody generation. These have shown early promise as plaque-directed in vivo targeting agents. Further scientific evaluation and reagent development remains before the practical utility of employing such reagents or methods in the care of patients with atherosclerosis-related diseases can be ascertained.

A preoperative chymopapain sensitivity test for chemonucleolysis candidates.

A fluorescence enzyme immunoassay (ChymoFAST test) for the quantitation of chymopapain-specific IgE antibody concentration in human serum is described. The IgE antibody is recognized as the major immunologic mediator for anaphylactic reactions. Serum chymopapain-specific IgE levels serve as an objective screening method for identifying patients most likely to tolerate chymopapain, thereby minimizing associated risks. Preoperative sera obtained from 11,658 chemonucleolysis candidates were tested, of which 0.94% (110/11,658) were found to be positive. Good predictive values for both positive and negative findings are evident based on 4776 postmarketing surveillances obtained from the surgeons.

Chymopapain-induced hypersensitivity following chemonucleolysis.

This study describes the changes in chymopapain-specific IgE antibody levels in patients following chemonucleolysis with Chymodiactin. Using the ChymoFAST method, chymopapain-specific IgE values were studied in 91 patients prior to and for 2 months post-Chymodiactin chemonucleolysis. A total of 8.8% (17/91) developed IgE levels greater than or equal to 0.06 IU/ml. Those patients with detectable IgE levels prior to chemonucleolysis were more likely than those with nondetectable preinjection levels (36.4% versus 4%) to develop chymopapain-specific IgE levels greater than or equal to 0.06 IU/ml.

Bacteria-specific IgE in patients with nasal polyposis. A preliminary report.

Sixty-one patients exhibiting chronic nasal polyposis had their conditions evaluated by radioallergosorbent tests to determine the presence of bacteria-specific serum IgE. Fifty-nine patients exhibited positive serum IgE to at least one of 11 bacteria tested. There are several implications of the collected data.