Resiquimod as an immunologic adjuvant for NY-ESO-1 protein vaccination in patients with high-risk melanoma.

The Toll-like receptor (TLR) 7/8 agonist resiquimod has been used as an immune adjuvant in cancer vaccines. We evaluated the safety and immunogenicity of the cancer testis antigen NY-ESO-1 given in combination with Montanide (Seppic) with or without resiquimod in patients with high-risk melanoma. In part I of the study, patients received 100 µg of full-length NY-ESO-1 protein emulsified in 1.25 mL of Montanide (day 1) followed by topical application of 1,000 mg of 0.2% resiquimod gel on days 1 and 3 (cohort 1) versus days 1, 3, and 5 (cohort 2) of a 21-day cycle. In part II, patients were randomized to receive 100-µg NY-ESO-1 protein plus Montanide (day 1) followed by topical application of placebo gel [(arm A; n = 8) or 1,000 mg of 0.2% resiquimod gel (arm B; n = 12)] using the dosing regimen established in part I. The vaccine regimens were generally well tolerated. NY-ESO-1-specific humoral responses were induced or boosted in all patients, many of whom had high titer antibodies. In part II, 16 of 20 patients in both arms had NY-ESO-1-specific CD4⁺ T-cell responses. CD8⁺ T-cell responses were only seen in 3 of 12 patients in arm B. Patients with TLR7 SNP rs179008 had a greater likelihood of developing NY-ESO-1-specific CD8⁺ responses. In conclusion, NY-ESO-1 protein in combination with Montanide with or without topical resiquimod is safe and induces both antibody and CD4⁺ T-cell responses in the majority of patients; the small proportion of CD8⁺ T-cell responses suggests that the addition of topical resiquimod to Montanide is not sufficient to induce consistent NY-ESO-1-specific CD8⁺ T-cell responses.

MAGE-specific T cells detected directly ex-vivo correlate with complete remission in metastatic breast cancer patients after sequential immune-endocrine therapy.

Studies suggest that conventional cancer therapies given after immunotherapy (IT) can boost antitumor immunity and possibly improve response rates and progression-free survival. We report two cases of metastatic breast cancer with durable complete responses (CRs) after sequential IT and endocrine therapy. Immune analyses of these long-term disease-free breast cancer patients previously treated with imiquimod (IMQ) suggest in-situ vaccination is achieved by topical application of the TLR-7 agonist directly onto tumors. Furthermore, IT-induced antigen-specific T cells were expanded by subsequent endocrine therapy and correlated with response, persisting > 2 years. Our findings therefore suggest that the induction/boosting of polyfunctional tumor antigen-specific T in response to sequential immune endocrine therapy and detected directly ex-vivo can serve as a peripheral blood biomarker for true clinical benefit.

Immunization of malignant melanoma patients with full-length NY-ESO-1 protein using TLR7 agonist imiquimod as vaccine adjuvant.

T cell-mediated immunity to microbes and to cancer can be enhanced by the activation of dendritic cells (DCs) via TLRs. In this study, we evaluated the safety and feasibility of topical imiquimod, a TLR7 agonist, in a series of vaccinations against the cancer/testis Ag NY-ESO-1 in patients with malignant melanoma. Recombinant, full-length NY-ESO-1 protein was administered intradermally into imiquimod preconditioned sites followed by additional topical applications of imiquimod. The regimen was very well tolerated with only mild and transient local reactions and constitutional symptoms. Secondarily, we examined the systemic immune response induced by the imiquimod/NY-ESO-1 combination, and show that it elicited both humoral and cellular responses in a significant fraction of patients. Skin biopsies were assessed for imiquimod's in situ immunomodulatory effects. Compared with untreated skin, topical imiquimod induced dermal mononuclear cell infiltrates in all patients composed primarily of T cells, monocytes, macrophages, myeloid DCs, NK cells, and, to a lesser extent, plasmacytoid DCs. DC activation was evident. This study demonstrates the feasibility and excellent safety profile of a topically applied TLR7 agonist used as a vaccine adjuvant in cancer patients. Imiquimod's adjuvant effects require further evaluation and likely need optimization of parameters such as formulation, dose, and timing relative to Ag exposure for maximal immunogenicity.

Vaccination with NY-ESO-1 protein and CpG in Montanide induces integrated antibody/Th1 responses and CD8 T cells through cross-priming.

The use of recombinant tumor antigen proteins is a realistic approach for the development of generic cancer vaccines, but the potential of this type of vaccines to induce specific CD8(+) T cell responses, through in vivo cross-priming, has remained unclear. In this article, we report that repeated vaccination of cancer patients with recombinant NY-ESO-1 protein, Montanide ISA-51, and CpG ODN 7909, a potent stimulator of B cells and T helper type 1 (Th1)-type immunity, resulted in the early induction of specific integrated CD4(+) Th cells and antibody responses in most vaccinated patients, followed by the development of later CD8(+) T cell responses in a fraction of them. The correlation between antibody and T cell responses, together with the ability of vaccine-induced antibodies to promote in vitro cross-presentation of NY-ESO-1 by dendritic cells to vaccine-induced CD8(+) T cells, indicated that elicitation of NY-ESO-1-specific CD8(+) T cell responses by cross-priming in vivo was associated with the induction of adequate levels of specific antibodies. Together, our data provide clear evidence of in vivo cross-priming of specific cytotoxic T lymphocytes by a recombinant tumor antigen vaccine, underline the importance of specific antibody induction for the cross-priming to occur, and support the use of this type of formulation for the further development of efficient cancer vaccines.

Clockwise and counterclockwise rotating shifts: effects on temperature and neuroendocrine measures.

INTRODUCTION: Most researchers suggest that shift rotation in a clockwise (CW) direction produces less disruption of circadian rhythms than rotations in a counterclockwise (CCW) direction. This is based on extrapolation from quasi-experimental studies of shift workers and research on the effects of jet lag that indicate that westward travel results in less disruption of circadian rhythms. METHODS: The effect of direction of rotation on cortisol, melatonin, and rectal temperature was examined in participants randomly assigned to either a CW (n = 14) or CCW (n = 14) shift rotation. RESULTS: Results revealed lower amplitude [F(1,21) = 14.6, p < 0.05] and a delay of the acrophase [F(1,21) = 4.4, p < 0.05] in temperature for the CCW group. Sample time effects for cortisol and melatonin revealed normal circadian variation for both rotations, although melatonin levels on the midnight shift were clearly suppressed relative to baseline melatonin levels. DISCUSSION: The changes observed in the temperature rhythm for the CCW rotation may be related to adaptation or shift-work hardiness; however, it is unclear if these differences indicate beneficial or detrimental changes for the individual. The fact that there were no significant effects of rotation condition for cortisol or melatonin argues against a detrimental effect. These findings are bolstered by performance and sleep data, reported elsewhere from this study, indicating a lack of significant differences as a function of rotation condition.

Clockwise and counterclockwise rotating shifts: effects on vigilance and performance.

INTRODUCTION: Arguments against counterclockwise shift schedules, such as those used in air traffic control, are prevalent in the literature; however, few studies have examined direction of rotation in rapidly rotating schedules. The present study directly compared clockwise (CW) and counterclockwise (CCW) rapidly rotating shiftwork schedules on measures of complex and vigilance task performance. METHODS: Participants (n = 28) worked day shifts for the first week of the study (0800-1600 h), followed by 2 wk of either a CW (n = 14) or CCW (n = 14) shiftwork schedule. Participants completed three 1.5-h sessions on the Multiple Task Performance Battery (MTPB) on each shift. Each session contained active- and passive-task components. In addition, participants completed a 0.5-h Bakan Vigilance Test at the beginning and end of each shift. RESULTS: A three-way, rotation condition by shift by session interaction (F (8,19) = 3.0, p < 0.05) for the active task composite scores and a rotation condition by shift interaction (F (4,23) = 6.2, p < 0.05) for the Bakan Vigilance Task indicated that effects of rotation condition were modulated by shift type, such that on particular shifts, performance in the CCW rotation was actually better than in the CW rotation. DISCUSSION: These data do not support the hypothesis that a CW rotation will result in better outcomes on complex or vigilance task performance. The results of this study indicate that two problem areas in both CW and CCW rapidly rotating shift schedules are early morning and midnight shifts.

Clockwise and counterclockwise rotating shifts: effects on sleep duration, timing, and quality.

INTRODUCTION: Many air traffic control specialists (ATCS) work relatively unique counterclockwise, rapidly rotating shift schedules. Researchers recommend, however, that if rotating schedules are to be used, they should rotate in a clockwise (CW), rather than a counterclockwise (CCW) direction. Unfortunately, few studies have examined CW and CCW rapidly rotating shifts. This study was designed to partially remedy this lack by examining the effects of both types of schedules on sleep duration, timing, and quality. METHODS: Participants (n = 28) worked a week of day shifts (0800-1600 h) followed by two weeks of either a CW (n = 14) or CCW (n = 14) shiftwork schedule, including early morning (0600-1400 h), afternoon (1400-2200 h), and midnight shifts (2200-0600 h). Participants recorded sleep data and subjective ratings of sleep quality, sleepiness, and mood in daily logbooks and wore wrist activity sensors to provide an objective source of sleep/wake data. RESULTS: There was no effect of rotation condition for any of the sleep measures. A main effect for Sleep Period (F (5,16) = 40.9, p < 0.05) indicated that both groups received less sleep prior to the early morning shifts than before the afternoon and midnight shifts. Sleepiness ratings were highest at the end of the midnight shift. DISCUSSION: Rotation condition did not play a significant role in the results of any of the sleep or subjective measures. The results of this study indicate that two problem areas in both CW and CCW rapidly rotating shift schedules are early morning and midnight shifts.