ABSTRACT
Adjuvanted protein vaccines offer high efficacy, yet most potent adjuvants remain proprietary. Several adjuvant compounds are being developed by the Vaccine Formulation Institute in Switzerland for global open access clinical use. In the context of the R21 malaria vaccine, in a mouse challenge model, we characterize the efficacy and mechanism of action of four Vaccine Formulation Institute adjuvants: two liposomal (LQ and LMQ) and two squalene emulsion-based adjuvants (SQ and SMQ), containing QS-21 saponin (Q) and optionally a synthetic TLR4 agonist (M). Two R21 vaccine formulations, R21/LMQ and R21/SQ, offer the highest protection (81%-100%), yet they trigger different innate sensing mechanisms in macrophages with LMQ, but not SQ, activating the NLRP3 inflammasome. The resulting in vivo adaptive responses have a different TH1/TH2 balance and engage divergent innate pathways while retaining high protective efficacy. We describe how modular changes in vaccine formulation allow for the dissection of the underlying immune pathways, enabling future mechanistically informed vaccine design.
Subject(s)
Malaria Vaccines , Malaria , Animals , Mice , Liposomes , Th1 Cells , Emulsions , Adjuvants, Immunologic/pharmacology , Malaria/prevention & controlABSTRACT
In the field of malaria vaccines, there are many barriers to moving lead candidates from the bench into developmental programmes before clinical testing. Many of the same challenges are to be found in the field of vaccines for other infectious diseases. Here, we briefly outline the process of pre-clinical development to help identify ways to support the translation of laboratory-based information into viable vaccine candidates.
Subject(s)
Antigens, Protozoan/immunology , Malaria Vaccines , Malaria/prevention & control , Clinical Trials as Topic , Humans , Malaria/parasitology , Malaria Vaccines/standards , Plasmodium/immunologySubject(s)
Forkhead Transcription Factors/genetics , Multiple Myeloma/genetics , Plasma Cells/metabolism , Repressor Proteins/genetics , Translocation, Genetic , Aged , Forkhead Transcription Factors/metabolism , Gene Expression Regulation, Neoplastic , Humans , Male , Multiple Myeloma/pathology , Repressor Proteins/metabolismABSTRACT
In view of the certain anatomic site-dependent frequency of chromosomal translocations involved in extranodal marginal zone B cell lymphoma of mucosa-associated lymphoid tissue (MALT lymphoma) pathogenesis, 17 salivary gland MALT lymphoma cases were analyzed for MALT1 and FOXP1 translocations. B cell CLL/lymphoma 10 (BCL10) and forkhead box PA (FOXP1) protein expression were studied by immunohistochemistry and translocations identified using fluorescence in situ hybridization (FISH)-specific probes FOXP1, t(11;18)(q21;q21)/API2-MALT1 and t(14;18)(q32;q21)/IgH-MALT1. None of the 11 analyzed cases showed FOXP1 rearrangement or amplification. The t(11;18) was present in five of 13 cases and the t(14;18) in three of 13 cases. MALT1 translocations were mostly mutually exclusive except in a single case. FOXP1 protein expression showed differences in the proportion of tumor cells with nuclear expression but not in their intensity, with the exception of one case where very intense nuclear staining was noted. BCL10 nuclear expression was present in four of 17 cases, two of which lacked t(11;18). Our results suggest that MALT1-specific translocations and FOXP1 rearrangements are not commonly involved in pathogenesis. A case with strong FOXP1 protein expression indicates the possibility that the upregulation of FOXP1 expression is significant in a small subset of salivary gland MALT lymphomas. Also a single case in which both MALT1 translocations were present indicates that these are not always mutually exclusive.
Subject(s)
Adaptor Proteins, Signal Transducing/metabolism , Caspases/metabolism , Forkhead Transcription Factors/metabolism , Lymphoma, B-Cell, Marginal Zone/metabolism , Neoplasm Proteins/metabolism , Repressor Proteins/metabolism , Salivary Gland Neoplasms/metabolism , Adaptor Proteins, Signal Transducing/genetics , B-Cell CLL-Lymphoma 10 Protein , Caspases/genetics , Forkhead Transcription Factors/genetics , Gene Expression Regulation, Neoplastic , Humans , In Situ Hybridization, Fluorescence , Lymphoma, B-Cell, Marginal Zone/etiology , Lymphoma, B-Cell, Marginal Zone/genetics , Lymphoma, B-Cell, Marginal Zone/pathology , Mucosa-Associated Lymphoid Tissue Lymphoma Translocation 1 Protein , Neoplasm Proteins/genetics , Repressor Proteins/genetics , Salivary Gland Neoplasms/etiology , Salivary Gland Neoplasms/genetics , Salivary Gland Neoplasms/pathology , Translocation, Genetic , Up-Regulation/geneticsABSTRACT
Over the last decade, fluorescence in situ hybridization (FISH) has become a firmly established technique in the diagnosis and assessment of lymphoid malignancies. However, this technique is not wide-ly used in the routine diagnostic evaluation of paraffin-embedded biopsies, most likely because of a perception that it is technically more demanding. There are also uncertainties regarding diagnostic thresholds and the way in which results should be interpreted. In this Review, we describe practical strategies for using FISH analysis to detect lymphoma-associated chromosomal abnormalities in routine paraffin-embedded lymphoma biopsies. Furthermore, we provide proposals on how FISH results should be interpreted (including how to calculate cutoff levels for FISH probes), recorded, and reported. An online appendix (available at http://jmd.amjpathol.org) details various simple, yet robust procedures for paraffin FISH analysis; it also provides additional information on the production of FISH probes, evaluating and reporting FISH results, sources for reagents and equipment, and troubleshooting. We hope that these suggestions will make FISH technology for the study of lymphoma biopsies more accessible to routine diagnostic and research laboratories.
