T cell-loaded injectable chitosan scaffold shows short-term efficacy in localised cancer immunotherapy in mice.

Adoptive cell therapy (ACT) shows success against treatment-resistant cancers, but is limited by the large number of intravenously delivered T cells required and toxicity related to systemic administration. In this work, we hypothesized that localized T cell delivery in an in situ gelling chitosan hydrogel will allow similar treatment efficacy despite delivering fewer cells than systemic intravenous delivery. A rapidly gelling chitosan gel with good mechanical properties was used for this study. Gel biocompatibility and biodegradability were tested over 8 weeks in mice. No adverse effects were observed. The gel elicited a local granulomatous reaction (foreign body reaction), degrading by about 75% volume at 8 weeks. The survival, escape and bioactivity against the tumour cells of encapsulated murine lymphocytes (OT-I) and human Jurkat cells were confirmed in vitro by live/dead assay and flow cytometry. Efficacy was studied using a mouse tumour model where the injected OT-I can specifically recognize and attack ovalbumin (OVA) protein-expressing tumours. The OT-I cell delivery scaffold was compared to untreated controls, OT-I in saline and intravenous systemic treatment with 3-fold more OT-I, observing tumour growth and localization by intravital microscopy and histology. Gel-encapsulated OT-I limited tumour growth significantly up to 11 days after treatment compared to that of untreated mice and mice with longer PBS-suspended OT-I treatment (9 days), but slightly less than that of mice with IV-delivered OT-I treatment (14 days). No significant difference was observed when directly comparing the gel and IV treatments. Although further optimization of the treatment is required, this work shows the feasibility and potential of the chitosan gel for localised OT-I delivery in cancer immunotherapy.

Quantitation of Coxsackievirus A21 Viral Proteins in Mixtures of Empty and Full Capsids Using Capillary Western.

A prototype strain of Coxsackievirus A21 (CVA21) is being evaluated as an oncolytic virus immunotherapy. CVA21 preferentially lyses cells that upregulate the expression of intercellular adhesion molecule 1, which includes some types of tumor cells. CVA21 has an icosahedral capsid structure made up of 60 protein subunits encapsidating a viral RNA genome with a particle diameter size of 30 nm. Rapid and robust analytical methods to quantify CVA21 total, empty, and full virus particles are important to support the process development, meet regulatory requirements, and validate manufacturing processes. In this study, we demonstrate the detection of all four CVA21 capsid proteins, VP1, VP2, VP3, and VP4, as well as VP0, a surrogate for empty particles, using in-house-generated antibodies. An automated and quantitative capillary Western blot assay, Simple Western, was developed using these antibodies to quantify CVA21 total particles through VP1, empty particles through VP0, relative ratio of empty to full particles through VP0 and VP4, and the absolute ratio of empty to total particles through VP0 and VP1. Finally, this Simple Western method was used to support CVA21 cell culture and purification process optimization as a high-throughput analytical tool to make rapid process decisions.

Biomaterials for enhanced immunotherapy.

Cancer immunotherapies have revolutionized the treatment of numerous cancers, with exciting results often superior to conventional treatments, such as surgery and chemotherapy. Despite this success, limitations such as limited treatment persistence and toxic side effects remain to be addressed to further improve treatment efficacy. Biomaterials offer numerous advantages in the concentration, localization and controlled release of drugs, cancer antigens, and immune cells in order to improve the efficacy of these immunotherapies. This review summarizes and highlights the most recent advances in the use of biomaterials for immunotherapies including drug delivery and cancer vaccines, with a particular focus on biomaterials for immune cell delivery.

Low-density lipoprotein receptor-related protein polymorphisms in patients with elevated factor VIII coagulant activity and venous thrombosis.

Elevated factor VIII coagulant activity (FVIII:C) levels (>150 IU/dl) represent a prevalent independent risk factor for venous thromboembolism (VTE). Low-density lipoprotein receptor-related protein (LRP) is involved in factor VIII clearance in vivo, and elevated FVIII:C was a feature of the LRP knockout mouse model. Three coding polymorphisms of LRP1 (exon 3, C766T; exon 14, A217V; and exon 39, D2080N), together with an insertion/deletion polymorphism within the first intron of lipoprotein receptor-associated protein (LRPAP1), have been identified. In addition, LRP1 2080D was recently reported to be associated with increased plasma FVIII:C levels in normal individuals. In this study, we investigated the role of these four polymorphisms in patients with objectively confirmed VTE and elevated FVIII:C levels. In our control group, genotype distributions were consistent with previous reports. Neither the allele frequencies nor genotype distributions at LRP1 A217V, LRP1 D2080N and LRPAP1 intron 1 were significantly different between the elevated FVIII:C and control groups. In contrast to previous reports, we found no effect of LRP1 D2080N genotype on plasma FVIII:C levels in normal individuals. More importantly, prevalence of the LRP1 2080D allele was not increased in the group of patients with high FVIII:C and VTE. We conclude that LRP1 and LRPAP1 polymorphisms are not responsible for high FVIII:C levels in patients with VTE.

Progress report on the development of child abuse prevention, identification, and treatment systems in Eastern Europe.

PROBLEM: After the Soviet Union dissolved in 1989, it became apparent that there was little recognition of the problems of child abuse and neglect, professionally, legally, or societally. There were no effective systems or laws in place to deal with these problems. METHOD: Beginning in 1995 the Children's Mental Health Alliance, in conjunction with the Open Society Institute began conducting trainings in Eastern Europe [Journal of the American Academy of Child Adolescent Psychiatry 39 (2000) 660]. Originally 18 countries from the Baltics to the Balkans participated. A program was elaborated which would proceed in several stages: (1) training mental health professionals to deal with child abuse and neglect (CAN); (2) teaching multidisciplinary team work and fostering the development of multidisciplinary NGOs focused on CAN; (3) promoting the self-sufficiency of these NGO's which would then facilitate social and legal reform and increase public awareness of the problem. Specific methods included multi-national trainings, assignment of mentors to the developing teams who maintained weekly contact with the teams and made yearly site visits to their countries, and overseeing project grants from OSI. RESULTS: NGO's had been established and registered in 11 countries, many establishing a network of programs within their countries. By 2000, over 3800 mental health professional had been trained, either directly by the program or by the trainees of the program. By the end of 2000, over 17,000 other professionals (lawyers, police, judges, educators, other physicians, etc.) had been trained by the network. CONCLUSION: While more work needs to be in this region, the teams in 11 countries have made solid starts.

Mother-daughter relationships and child sexual abuse: a pilot study of 35 dyads.

In a pilot study, the authors examine features of mothers' relationships to their mothers, spouses, and daughters associated with their daughters' male-perpetrated child sexual abuse. Fifteen inner-city Latino mothers and daughters (ages 1-9 years), referred by child protective agencies for highly suspect or confirmed child sexual abuse, were compared to 20 matched control dyads. Significantly more case mothers than controls reported relational disturbances intergenerationally, including hostility toward their daughters. Mother-daughter relationships should be thoroughly assessed when evaluating a child for child sexual abuse.