Active specific immunotherapy of malignant melanoma with anti-idiotypic monoclonal antibodies

In this study, we have utilized the human high molecular weight-melanoma associated antigen (HMW-MAA) as a target for active specific immunotherapy with mouse anti-idiotypic (anti-id) monoclonal antibodies (mAb) in patients with malignant melanoma. After having summarized the characteristics of HMW-MAA which account for its selection as a target for immunotherapy, we describe the development and characterization of mouse anti-id mAb MK2-23 which bears the internal image of HMW-MAA. Furthermore, we describe the results of the first clinical trial performed with mouse anti-id mAb MK2-23 in patients with malignant melanoma.     

Surgical adjuvant active specific immunotherapy for patients with stage III melanoma: the final analysis of data from a phase III, randomized, double-blind, multicenter vaccinia melanoma oncolysate trial

BACKGROUND: A phase III, randomized, double-blind, multicenter trial of active specific immunotherapy (ASI) using vaccinia melanoma oncolysate (VMO) was performed in patients with stage III (American Joint Commission on Cancer) melanoma to determine the efficacy of VMO to increase the disease-free interval (DFI) or overall survival (OS) in these patients. Two interim analyses of data from this trial were performed in May 1994 and June 1995. Although the results from these analyses showed no statistically significant improvement in DFI or OS in all patients using VMO, two subsets-men aged 44-57 years with one to five positive nodes and all patients with clinical stage I and pathologic stage II disease-showed an overall survival advantage with VMO therapy. A final analysis of data from this trial was performed in May 1996 and is reported here. The design of future melanoma vaccine trials is discussed based on information learned from this first randomized, multicenter trial of ASI therapy. STUDY DESIGN: A polyvalent VMO was prepared using melanoma cells derived from four melanoma cell lines and vaccinia vaccine virus (V). Patients were accrued from 11 United States institutions and were randomized by the Statistical Center at the University of Alabama, Birmingham. Two hundred fifty patients were randomized to treatment with either VMO (1 U containing 2 mg of total protein derived from 5 x 10(6) melanoma cells and 10(5.6) 50% tissue culture infectious dose of vaccinia virus) or control V (1 U containing 10(5.4) 50% tissue culture infectious dose of vaccinia virus) once a week for 13 weeks and then once every 2 weeks for a total of 12 months, or until recurrence. Patient data were collected by the Statistical Center and analyzed as of May 1996 for DFI and OS using Wilcoxon test and log-rank analysis. RESULTS: Two hundred seventeen patients were found to be eligible according to the inclusion criteria. Data from these patients were analyzed for DFI and OS after a median followup of 46.3 months (50.2 months for VMO and 41.3 months for V). This final analysis showed no statistically significant increase in either DFI (p = 0.61) or OS (p = 0.79) of patients treated with VMO (n = 104) compared with V (n = 113). At 2-, 3-, and 5-year intervals, 47.8%, 43.8%, and 41.7% of patients treated with VMO were disease-free, respectively, compared with 51.2%, 44.8%, and 40.4% of patients treated with V. At the same intervals, 70.0%, 60.0%, and 48.6% of patients treated with VMO survived, compared with 65.4%, 55.6%, and 48.2% of patients treated with V. In a retrospective subset analysis, male patients aged 44-57 years (n = 20) with one to five positive nodes showed 18.9%, 26.82%, and 21.3% improvement in survival at 2-, 3-, and 5-year intervals, respectively, after treatment with VMO when compared with V (n = 18) (p = 0.046). CONCLUSIONS: This study was a randomized, multicenter, placebo-controlled evaluation of an active specific immunotherapeutic agent to increase the DFI or OS of patients with stage III melanoma in a surgical adjuvant setting. In this trial, ASI with VMO when compared with V showed no difference in either DFI or OS. In a retrospective subset analysis, however, a subset of men with one to five positive nodes, between the ages of 44 and 57 years, showed a survival advantage with VMO. This result suggests that one must include a detailed subset analysis in the design of future trials of ASI for patients with American Joint Commission on Cancer stage III melanoma. An appropriate control arm also must be included in ASI trials.     

A novel tumor-specific human single-chain Fv selected from an active specific immunotherapy phage display library

The nucleus accumbens (NAcc) can be subdivided into 'core' and 'shell' based on anatomical connections and histochemical markers. Previous studies have demonstrated dopamine-beta-hydroxylase immunoreactive (DBH-ir) fibers in the NAcc shell, but the source of these noradrenergic (NE) afferents has not been determined. Therefore, we have investigated in detail the anatomy of NE afferents to this subregion. Dual immunohistochemistry for DBH and substance P demonstrated numerous DBH-ir fibers in the caudal NAcc shell. Neurons projecting to the NAcc were identified with Fluoro-Gold (FG) or cholera toxin B (CTb) retrograde tracing and tyrosine hydroxylase (TH) immunohistochemistry. Single- and double-labeled neurons were observed in the A2 and A1 NE cell groups following FG injections into the caudal NAcc shell. Numerous FG and CTb single-labeled neurons were found in the rostral locus coeruleus (LC), subcoeruleus and pericoerulear dendritic region, with an occasional double-labeled neuron in the LC. Few labeled neurons were seen in the brainstem after FG injections into the NAcc core, consistent with the lack of DBH-ir in this subterritory. To confirm these results, injections of Phaseolus vulgaris leucoagglutinin or biotinylated dextran amine were made into the LC or nucleus tractus solitarius (NTS). Virtually no labeled fibers were observed in the NAcc following injections into central LC. However, fibers were observed in the NAcc shell after injections in the NTS. These results indicate that the primary source(s) of NE afferents to the NAcc shell is the A2 region of the NTS, with lesser contributions from A1 and LC.     

Adenovirus-mediated expression of melanoma antigen gp75 as immunotherapy for metastatic melanoma

Melanocyte differentiation antigens, such as the brown locus protein gp75, are potential biological targets for immunotherapy. We investigated whether expression of the murine gp75 cDNA mediated by an adenovirus (Ad) vector could induce melanoma rejection using this model self antigen that usually induces tolerance, and whether Ad vector-directed production of interleukin-2 (IL2) might augment this response. To evaluate this approach, Ad vectors were constructed containing the murine gp75 cDNA (Ad.gp75) and the human IL2 cDNA (Ad.IL2). Efficacy was evaluated in C57BI/6 mice challenged i.v. with 10(5) B16 cells, using the number of lung metastases as the efficacy parameter. Naive control mice developed 175 +/- 12 metastases by day 14. Controls receiving intranasal Ad.IL2 1 day after B16 cell injection, intraperitoneal (i.p.) mitomycin-C-treated B16 cells +/- i.p. Ad.IL2 before B16 cell challenge and Ad.beta gal-treated mice had similar numbers of metastases as controls (P > 0.1). In marked contrast, preimmunization with intradermal Ad.gp75 provided dramatic reduction in the number of lung metastases (52 +/- 7, 29% of control). Addition of regional (intranasal delivery to the lung) Ad.IL2 to intradermal Ad.gp75 preimmunization 1 day following tumor challenge provided further protection (18 +/- 6, 10% of control). Depletion of CD4+ and CD8+ T-cell subsets effectively blocked the protective effect seen following immunization. Adoptive transfer of macrophage-depleted splenocytes from Ad.gp75-immunized mice similarly afforded significant protection against B16 tumor cell challenge. Further, serum obtained 21 days following Ad.gp75 immunization showed no detectable anti-gp75 antibody by immunoprecipitation. These results suggest that immunization with Ad.gp75 induces cellular immune responses that are capable of rejecting B16 melanoma in a host that is usually tolerant to gp75 antigen.     

Specific immunotherapy using allergens apropos of specific immunotherapy by the sublingual route

The present number of publications an allergen specific immunotherapy (ITa) by clinically controlled tests, double blind, is objective proof of the value of this treatment in the allergy pathologies mediated by IgE. The mechanisms of action of Ita are better understood, taking account of the intervention of TH2 and TH1 cells with the respective involvement of cytokines IL4-IL5, favouring the synthesis of IgE and interferon gamma (IFN gamma) slowing the synthesis of IgE. Adjuvants of bacterial origin influence the balance TH2/TH1 by blocking the production of TH2 cells. Ita by the sublingual route (SLIT) has been the objective of double blind clinical studies, with favourable results and good tolerance, of a number of allergens, pollens (grass, ragweed, parietaria, olive) and D. pteronyssinus mites. Application of this technique of SLIT to foods and drugs reveals more a mechanism of customisation than induction of cellular acticity, of a mucosal secretory IgA and an oral tolerance with systemic response, mechanisms that are involved in SLIT. Taking into account the restrictions of ITa, SLIT allows inclusion of other allergic patients, such as children receiving injections and subjects who are not inclined to observe and comply with treatment.... The different techniques used for classical ITa by the subcutaneous route are also useable for SLIT with a very good tolerance, apart from some transitory digestive problems. SLIT is no longer a technique of the future, but already one of the present time, currently used and which gives a renewal of value in the form of therapy that is ITa for IgE-mediated allergy pathology.     

Active immunotherapy with allogeneic tumor cell vaccines: present status

This review will concentrate on allogeneic vaccines for melanoma The important principles of melanoma vaccine effectiveness are discussed in detail, followed by a review of the progress of several clinical trials investigating allogeneic vaccines. No therapeutic cancer vaccine has yet been approved for general use by the US Food and Drug Administration. However, much progress has been made in the field of vaccine immunotherapy, especially for the treatment of melanoma. Active immunotherapy with tumor vaccines is progressing rapidly as an emerging option for cancer therapy.     

Mechanisms of specific immunotherapy of allergic diseases

A government agency and its contractors employing nearly 96,000 workers throughout the country was surveyed for documented incidents of violence in the workplace. Thirty-five occupational medicine and related professionals (36% of those surveyed) from 27 locations returned the questionnaire. Of the responders, 20 individuals reported 74 incidents of workplace violence, with nearly 30% of these occurrences involving weapons, including 11 with guns. In a companion survey of human resource departments from 28 locations, there were 16 responders (57% of those surveyed) with 13 of them documenting 96 additional incidents. No duplicate reporting of the same event occurred between the two surveys. Approximately 70% of the agency workers were employed in locations covered by the 51 responders. Although the data are limited, the number of incidents and level of violence appear to be increasing over time. Of the 108 incidents for which time of occurrence was known, 32 were defined as "very serious," which included physical assault, threat or assault with a weapon, murder, suicide, or stalking. Verbal threats, verbal assaults, and vandalism were defined as "serious" incidents. A Cochran-Armitage trend test for an increasing proportion over time of "very serious" vs. "serious" events was statistically significant, with a P-value of 0.026.     

Efficacy of anti-intercellular adhesion molecule-1 immunotherapy on immune responses to allogeneic hepatocytes in mice

Adhesion molecules appear to play important roles in vascularized organ allograft rejection, because antibodies directed against them are effective in prolonging survival of vascularized organ allografts in rodents. However, the efficacy of these agents for cellular allografts is unknown. The current studies were undertaken to determine the role of intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) on host immune responses to purified hepatocytes. Host mice (C3H, H-2(k)) grafted with hepatocytes in sponge matrix allografts (HC-SMA) received IgG isotype control, anti-ICAM-1, or anti-VCAM-1 monoclonal antibody (mAb) on days 0 through 9 after grafting. Twelve to 14 days later, host cells infiltrating the HC-SMA were assessed for the development of allospecific cytolytic T cells (allo-CTLs). Treatment with anti-ICAM-1 or anti-VCAM-1 mAb resulted in significantly decreased recruitment of host cells into HC-SMA (P < .035). However, only anti-ICAM-1 mAb resulted in abrogation of development of allo-CTLs in HC-SMA (P = .001). C3H (H-2(k)) hosts grafted with allogeneic hepatocytes from control C57BL/6 (H-2(b)) or ICAM-1 knockout [H-2(b)] mice elicited the development of allo-CTLs in HC-SMA (P = not significant). Furthermore, there was no difference in the development of allo-CTLs in HC-SMA of control hosts [C57BL/6, H-2(b)] compared with ICAM-1 knockout hosts (H-2(b)) (P = not significant). Treatment with anti-ICAM-1 mAb had no effect on the development of allo-CTLs in ICAM-1 knockout (H-2(b)) hosts bearing HC-SMA. The immunosuppressive effect of host treatment with anti-ICAM-1 mAb does not appear to be a consequence of simple blockage of donor hepatocyte or host immune cell expression of ICAM-1, but suggests a potential inhibitory effect on host immune cell activation or function, as well as an effect on recruitment of host cells to the allograft.     

Active immunotherapy with ultraviolet B-irradiated autologous whole melanoma cells plus DETOX in patients with metastatic melanoma

Advances in transdermal therapeutic systems (TTS) and the technology involved have been rapid because of the sophistications of physiology and biology on skin, enhancing methodology for the skin penetration of drugs, and material and polymer sciences on the pharmaceutical additives. In this review paper, such progress was introduced and the future on the TTS was augured.     

Results of specific immunotherapy with Allergovit in patients with pollen allergy

The immunotherapy among patients with pollen allergy was performed by individuals composed Allergovit. The effects of this treatment was compared to patients with pollenosis who don't have desensitization because they would'nt it. This results indicated that is need to do the nasal provocation test before the choice of antigens for the specific immunotherapy because the prick skin test was very often positive but patients don't have the symptoms. Precise composition of the vaccine improved the course in first season in 77.9 per cent, in second season in 87.5 per cent and in third season in 91.6 per cent.     

Immunological reactions of skin melanoma patients to nonspecific and adaptive immunotherapy

Patients with malignant skin melanoma were given different kinds of immunotherapy; nonspecific immunotherapy with BCG vaccine, adaptive immunotherapy with methotrexate or phytohemagglutinin activated autolymphocytes, and also a combination of polychemotherapy with BCG. The state of cell immunity was determined prior to, during and at the end of the therapy, in accordance with the courses. As a result of the conducted therapy an increased level of cell immunity was observed, that usually corresponded to clinical development of the disease. At the same time, the increase in immune response indices due to continuous administration of BCG vaccine is not related with the clinical course, while the persistent anergy or reduced level of cell immunity during this kind of therapy is a poor prognostic sign.     

Diagnostic problems before specific immunotherapy in patients with late-summer seasonal allergic rhinitis

Skin prick tests were performed on 245 patients with late summer seasonal allergic rhinitis, and of these patients, 135 specific serum IgE test were performed. On the basis of skin prick test results, 94% of the patients were found to be sensitive to Ragweed: 18% of these patients had monosensitisation to Ragweed, and 56% were sensitive not only to Ragweed but also to Mugwort. The correlation between results of skin prick tests and specific serum IgE tests was found to be very good (95%) with Ragweed antigen experiencing no problem in the diagnostic process before immunotherapy. However, in 48% of patients with positive skin prick tests to Mugwort the specific serum IgE was found to be negative. Before immunotherapy, a specific nasal provocation test was performed on 12 of these patients with Mugwort to examine the real sensitivity of the shock-organ. This careful allergen research will demonstrate which components of allergen extract should be used for immunotherapy in late summer seasonal allergic rhinitis patients.     

T-cell receptor beta variable region diversity in melanoma metastases after interleukin 2-based immunotherapy

Limited T-cell receptor (TCR) repertoire of tumor-infiltrating lymphocytes has been found in melanoma metastases and spontaneously regressing melanoma. Immunotherapy with INF-alpha/interleukin 2 can induce tumor regression in a proportion of patients with metastatic melanoma. We analyzed the gene expression of the TCR-beta variable (Vbeta) region of tumor-infiltrating lymphocytes from 16 melanoma metastases by subgroup-specific semiquantitative RNA PCR to investigate the influence of immunotherapy on the TCR pattern. In five progressing metastases before or after immunotherapy, no overexpression of Vbeta gene families was detectable, whereas in seven of seven metastases responding to IFN-alpha/interleukin 2 one to four Vbeta gene families were overexpressed. Preferential usage of certain Vbeta gene subgroups in patients sharing the same HLA class I molecules suggests a T-cell response to dominant public epitopes. Analysis of multiple specimens from the same patients gives evidence that this strong oligoclonal T-cell selection is induced or at least augmented by immunotherapy, supporting the functional relevance of this finding.     

Value of selected parameters for monitoring efficacy of specific immunotherapy in allergic disorders

Prevention, pharmacotherapy and specific immunotherapy are used as methods to treat patients with allergic disorders of respiratory system. SIT as a method of controlled supply of allergen, releases immunological mechanisms related to humoral, cellular and effector organic response. Role of those mechanisms and type of changes, taking place can be monitored with given immunological parameters. Most convincing proof of effectiveness of desensitisation, is diminishing of manifestation of clinical symptoms. In patients treated with obvious therapeutical progress, tolerance of increased dose of allergen was noted.