Vaccine and antibody-directed T cell tumour immunotherapy.

Clearer evidence for immune surveillance in malignancy and the identification of many new tumour-associated antigens (TAAs) have driven novel vaccine and antibody-targeted responses for therapy in cancer. The exploitation of active immunisation may be particularly favourable for TAA where tolerance is incomplete but passive immunisation may offer an additional strategy where the immune repertoire is affected by either tolerance or immune suppression. This review will consider how to utilise both active and passive types of therapy delivered by T cells in the context of the failure of tumour-specific immunity by presenting cancer patients. This article will outline the progress, problems and prospects of several different vaccine and antibody-targeted approaches for immunotherapy of cancer where proof of principle pre-clinical studies have been or will soon be translated into the clinic. Two examples of vaccination-based therapies where both T cell- and antibody-mediated anti-tumour responses are likely to be relevant and two examples of oncofoetal antigen-specific antibody-directed T cell therapies are described in the following sections: (1) therapeutic vaccination against human papillomavirus (HPV) antigens in cervical neoplasia; (2) B cell lymphoma vaccines including against immunoglobulin idiotype; (3) oncofoetal antigens as tumour targets for redirecting T cells with antibody strategies.

Immunological responses in women with human papillomavirus type 16 (HPV-16)-associated anogenital intraepithelial neoplasia induced by heterologous prime-boost HPV-16 oncogene vaccination.

PURPOSE: The purpose is to study the immunogenicity of heterologous prime-boost human papillomavirus (HPV) oncogene vaccination in patients with anogenital intraepithelial neoplasia (AGIN). EXPERIMENTAL DESIGN: Twenty-nine women with high-grade AGIN received three i.m. doses of TA-CIN (HPV-16 L2/E6/E7 protein) at four weekly intervals followed by a single dermal scarification of vaccinia HPV-16/18 E6/E7 and were followed up for 12 weeks. Immunity to HPV-16 was assessed by lymphoproliferation, IFN-gamma enzyme-linked immunospot (ELISPOT), and ELISA. RESULTS: The patient group significantly responded to TA-CIN and not to the control antigen HPV-6 L2/E7 at all postvaccination time points when compared with baseline responses (P < or = 0.05). Ten of the patients showed at least a 3-fold increase in TA-CIN-specific proliferation at one or more time points after vaccination. Comparison of stimulation with HPV-16 E6- or E7-GST fusion proteins showed that proliferative responses were biased to HPV-16 E6. This bias was also seen by IFN-gamma ELISPOT using overlapping peptides, with HPV-16 E6- or E7-specific T cells being detected in 9 and 2 patients, respectively. In addition, vaccination resulted in the induction of antibodies against the HPV-16 oncoproteins. Of the 6 clinical responders, 2 patients showed both a proliferative TA-CIN-specific response and an E6-specific IFN-gamma response, whereas 3 other patients displayed E6-specific reactivity only. Stable disease was recorded in 19 patients, 8 of whom showed a concomitant TA-CIN-specific proliferative and/or E6-specific T-cell response. Of the 4 progressors, 2 failed to make a T-cell response and 2 responded by either proliferation or E6 ELISPOT alone. CONCLUSIONS: The prime-boost regimen is immunogenic in AGIN patients (humoral and cellular immunity), but there is no simple relationship between induction of systemic HPV-16-specific immunity and clinical outcome. Other factors that may play a role in the eradication of long-term established AGIN lesions need to be determined to identify the patient group that would benefit from immunotherapy with the vaccines used in this study.

Immunological and clinical responses in women with vulval intraepithelial neoplasia vaccinated with a vaccinia virus encoding human papillomavirus 16/18 oncoproteins.

This study assessed the immunological and clinical responses of women with human papillomavirus (HPV) 16-associated high-grade vulval intraepithelial neoplasia (VIN) vaccinated with TA-HPV, a recombinant vaccinia virus encoding modified HPV 16 and 18 E6 and E7. Eighteen women with HPV 16-positive high-grade VIN were vaccinated with TA-HPV. The extent of their baseline disease was compared after 24 weeks by lesion measurements and histological analysis. Viral load was assessed pre- and postvaccination by real time PCR. Cell-mediated immunity to HPV 16 E6 and/or E7 peptides (HLA-A2 epitopes) or vaccinia-infected cell lysates was determined by IFN-gamma enzyme-linked immunospot (ELISPOT) and T cell proliferation using an HPV 16 L2E6E7 fusion protein. Antibodies were measured by ELISA using vaccinia-infected cell lysates or HPV 16 and 18 E6 and E7 glutathione S-transferase-fusion proteins. Lesion-infiltrating CD4(+), CD8(+), CD1a(+), and CD68(+) immune cells were assessed by immunohistochemistry. The single vaccination with TA-HPV was well tolerated, and all patients showed an increased ELISPOT and/or antibody response to vaccinia. There were significant differences in HPV-16 E7-specific ELISPOT and L2E6E7 proliferative responses in the patients at one or more time points postvaccination as compared with the prevaccination status; two patients showed transient increased antibody responses. Overall, 13 women showed an increased HPV 16-specific immune response by one or more methodologies after immunization. Eight patients demonstrated a reduction in lesion diameter of at least 50% and a further four patients showed significant symptom relief. Viral load was reduced or cleared in six of eight lesion responders but also in six of ten nonresponders. Before vaccination, clinical responders had significantly higher levels of lesion-associated CD4(+), CD8(+), and CD1a(+)-immune cells than nonresponders. There were no differences in CD68 (macrophages) between responders and nonresponders before or after vaccination. Nonresponders did show a significant increase in CD4(+)- and CD8(+)- but not CD1a(+)-immune cells postvaccination but at lower levels overall than responder patients. Local immune infiltration may be a critical factor in potential responsiveness to vaccine therapy in HPV-associated neoplasia and should be carefully monitored in future placebo-controlled trials of immunotherapy for VIN.

Human papillomavirus type 16 E2- and L1-specific serological and T-cell responses in women with vulval intraepithelial neoplasia.

Human papillomavirus type 16 (HPV-16)-associated vulval intraepithelial neoplasia (VIN) is frequently a chronic, multifocal high-grade condition with an appreciable risk of progression to vulval cancer. The requirement to treat women with VIN has recently stimulated the use of immunotherapy with E6/E7 oncogene vaccines. Animal models have shown that E2 may also be a useful vaccine target for HPV-associated disease; however, little is known about E2 immunity in humans. This study investigated the prevalence of HPV-16 E2-specific serological and T-cell responses in 18 women with HPV-16-associated VIN and 17 healthy volunteers. E2 responses were determined by full-length E2-GST ELISA with ELISPOT and proliferation assays using E2 C-terminal protein. As positive controls, HPV-16 L1 responses were measured using virus-like particles (VLPs) and L1-GST ELISA with ELISPOT and proliferation using VLPs as antigen. The VIN patients all showed a strong serological response to L1 compared with the healthy volunteers by VLP (15/18 vs 1/17, P<0.001) and L1-GST ELISA (18/18 vs 1/17, P<0.001). In contrast, L1-specific cellular immune responses were detected in a significant proportion of controls but were more prevalent in the VIN patients by proliferation assay (9/17 vs 17/18, P<0.02) and interferon-gamma ELISPOT (9/17 vs 13/18, P=not significant). Similar and low numbers of patients and controls were seropositive for E2-specific Ig (2/18 vs 1/17). In spite of previous studies showing the immunogenicity of E2 in eliciting primary T-cell responses in vitro, there was a low prevalence of E2 responses in the VIN patients and controls (2/18 vs 0/17).

Association between human leukocyte antigen polymorphism and human papillomavirus 16-positive vulval intraepithelial neoplasia in British women.

Polymorphisms in human leukocyte antigen (HLA) genes have been implicated in the risk for developing human papillomavirus (HPV)-associated cervical neoplasia. By comparison with local cadaver controls typed for HLA class I (n = 946) and II (n = 144) antigens, HPV-16-positive high grade vulval intraepithelial neoplasia patients (n = 42) showed significantly different frequencies of HLA-A2 [odds ratio (OR), 2.1; confidence interval (CI), 1.4-3.9], HLA-B7 (OR, 2.6; CI, 1.4-4.7), HLA-DRB1*01(01/02/04) (OR, 0.1; CI, 0.03-0.5), HLA-DRB1*11 (OR, 3.3; CI, 1.4-7.1), HLA-DRB1*13 (OR, 0), HLA-DQB1*05 (OR, 0.2; CI, 0.05-0.6), and HLA-DQB1*03032 (OR, 4.6; CI, 1.5-14.0). With the exception of HLA-B7 and HLA-DRB1*11, these significant differences were also seen comparative to local HPV-16-positive cervical carcinoma patients (n = 114), suggesting a specific immunogenetic contribution that is independent of HPV-16 infection in high-grade vulval intraepithelial neoplasia. Such factors are important to the development of HPV vaccines for treatment of cervical and vulval neoplasia.

Maternal serum activin, inhibin, human chorionic gonadotrophin and alpha-fetoprotein as second trimester predictors of pre-eclampsia.

OBJECTIVE: To compare the serum levels of human chorionic gonadotrophin (hCG), alpha-fetoprotein, activin A, inhibin A and inhibin isoforms containing pro and alphaC in the second trimester serum of women who subsequently developed hypertensive disorders of pregnancy with those who remained normotensive throughout pregnancy. DESIGN: Retrospective case-control study of 15-20 week serum samples matched for duration of storage at -20 degrees C. SETTING: Antenatal clinics at a teaching hospital in Scotland. SAMPLE: Second trimester serum samples of 39 women who subsequently developed pre-eclampsia, 31 who subsequently developed pregnancy-induced hypertension and 155 women who remained normotensive throughout pregnancy. MAIN OUTCOME MEASURES: hCG, alpha-fetoprotein, activin A, inhibin A and inhibin pro-alphaC serum levels. RESULTS: Activin A levels in serum were significantly elevated in women who later developed pregnancy-induced hypertension (26% increase compared with controls) and hCG levels were significantly elevated in women who later developed pre-eclampsia (24% increase compared with controls). alpha-Fetoprotein, inhibin A and inhibin pro-alphaC levels were not significantly elevated in the patient groups compared with their controls. CONCLUSIONS: A combination of analyses including second trimester serum activin A and hCG may yet prove to be helpful predictors of women at risk of hypertensive disorders of pregnancy. While the results proved significant, the effects reported in this study are too modest compared with natural variability to be useful as screening tools on their own.

The natural history of HPV infection of the uterine cervix. Long-term observational and histological data.

BACKGROUND: To investigate the natural history of HPV infection of the uterine cervix following a 10-year prospective observational study. PATIENTS AND METHODS: Three hundred and thirty women with cytological and colposcopic features of HPV infection +/- CIN 1 were recruited into the study. These women were followed up for a period of 10 years, during which time they had repeated colposcopic assessments and cytological monitoring Those women with evidence of high-grade disease at any stage of follow-up were treated by Large Loop excision of the transformation zone (LLETZ) and excluded from the study. At the end of the surveillance period, 179 women with evidence of persistent HPV infection alone or in conjunction with low-grade disease underwent a shallow diagnostic LLETZ. In total, 51 women defaulted from follow-up. RESULTS: Thirty-three per cent of the women with cytological and colposcopic features suggestive of persistent HPV infection alone during the 10-year follow-up period had histological evidence of CIN at the end of the study. Two thirds of this disease was graded as CIN 2 or 3. Nineteen per cent of the women with cytological and colposcopic features suggestive of persistent HPV infection and CIN 1 actually had high-grade CIN at histology (CIN 2 and 3). Only 6.1% of the study population had normal histological findings. CONCLUSION: The finding that a relatively large percentage of high-grade lesions might have been masked for anything up to a decade by a low-grade phenotype, coupled with the worrying percentage of defaulters from follow-up, indicate that an early decision for intervention (diagnostic and/or therapeutic) may be appropriate if a low-grade abnormality persists after a short surveillance period.

The use of vaccines in the prevention and treatment of cervical cancer.

The close association between high risk HPV infection and cervical carcinoma has provided the impetus for the development of prophylactic and therapeutic vaccination schedules. An effective prophylactic vaccine would obviate the need for population-based cervical screening programmes, while therapeutic vaccination might provide an effective adjunct to or replacement for conventional treatment for benign and malignant cervical disease. While the challenges associated with the design and implemention of immunotherapies are numerous, optimism remains high and it is expected that the next few decades will witness a revolutionary change in the way we treat cervical cancer and its premalignant lesions. A papillomavirus vaccine that prevented HPV infection on the one hand and acted against established disease on the other, would have a profound impact on one of the major cancers affecting women globally.

Bleeding after loop electrosurgical excision procedure performed in either the follicular or luteal phase of the menstrual cycle: a randomized trial.

OBJECTIVE: To estimate the perioperative or postoperative bleeding rates after treatment of cervical intraepithelial neoplasia by loop electrosurgical excision procedure in either the follicular or luteal phase of the menstrual cycle. METHODS: A randomized controlled trial was carried out to compare the outcomes in terms of primary and secondary hemorrhage between patients treated by loop electrosurgical excision procedure during either the follicular (30 women) or luteal phase (30 women) of the menstrual cycle. The two groups did not differ in terms of mean age, grade of cervical intraepithelial neoplasia, depth of excision, parity, and duration of menses. Primary outcome measures included the objective and subjective assessment of intraoperative and postoperative bleeding. RESULTS: Women treated during the luteal phase of the menstrual cycle experienced significantly more postoperative bleeding than women treated during the follicular phase, as assessed by the fall in hematocrit levels (P <.001) and subjective reports. Intraoperative bleeding was judged to be more severe in women treated during the luteal phase of the cycle by a single, blinded colposcopist (P =.02). These women also experienced higher levels of anxiety postoperatively, which resulted in more consultations with medical staff (P =.007). CONCLUSION: The use of loop electrosurgical excision procedure to treat cervical intraepithelial neoplasia results in less bleeding if performed during the follicular phase of the menstrual cycle.

Selective processing of weight- and shape-related words in bulimia nervosa: use of a computerised Stroop test.

A computerised Stroop colour-naming task was used to measure concerns about weight, shape, and eating in bulimia nervosa. Two versions of the computerised Stroop were compared, a voice-activated and a button-pressing-activated programme. Bulimia nervosa patients were significantly slower in colour naming shape- and weight-related words than their female age-matched controls. The button-pressing computerised Stroop was both more sensitive and more accurate at measuring colour-naming speeds than the voice-activated version. When the bulimia nervosa group were divided according to their Eating Attitudes Test (EAT) scores, those who showed extreme pathological attitudes to weight and shape were significantly slower in colour naming size words and in food disruption scores than those with a lesser degree of psychopathology. The computerised Stroop might be useful as a diagnostic tool and in the assessment of the effectiveness of therapy for the individual patient.

Management of ectopic pregnancy in a Scottish teaching hospital: implications for training.

An audit was undertaken to assess the management of 50 consecutive women with an ectopic pregnancy at a teaching hospital in Scotland. The ectopic pregnancy was removed laparoscopically in 62% and of these women 80% were discharged home on the first postoperative day. In a minority of patients, elements of substandard care were identified including failure to operate when the ectopic pregnancy had been visualised on ultrasound or in the presence of static hCG levels in patients with an empty uterus and adnexal pain. Fewer than 50% of consultants and no junior registrar reported competency in the laparoscopic management of these patients. We suggest that the management of ectopic pregnancy could be improved by following published algorithms with high diagnostic accuracy. In addition, either skilled endoscopic teams should be developed to take on the responsibility of the management of these patients or universal consultant competency in laparascopic surgery is required if we are to offer laparoscopy to all women with ectopic pregnancy.