Table of Contents

Abstract: To expand the open access publishing project of our newly founded company MDPI [1,2] based in Basel, Switzerland, we are in the process of launching new journals. Based on our success in running journals that represent key areas in science and technology, such as Molecules [3], Sensors [4], Energies [5], Viruses [6], Pharmaceuticals [7], Cancers [8] and Toxins [9], we are launching a new journal entitled Cells. It is an open access journal combining cell biology, molecular biology and biophysics, toward an understanding of cell structure, function and interactions. [...]

Abstract: I am honored to introduce the new journal Cells, which has been created to serve as a hub for disseminating new findings and discoveries in cell biology to researchers worldwide. Cells is an international, peer-reviewed, open-access journal on cell biology, molecular biology, and biophysics. Much has been accomplished in molecular and cell biology in the past thirty years. We have unraveled the structure of many proteins, nucleic acids and lipids, and learned much about cell structures, including lysosomes, mitochondria, cytoskeletal elements, cell membranes, endoplasmic reticulum, and nuclei. Now, it is time to apply this knowledge to answering the age-old question: “How do cells work?” It appears there is a strong demand for a journal focused on various functional aspects of cell biology, including such fundamental ones as gametogenesis, embryonic development, tissue regeneration, tumorigenesis, and aging. [...]

Abstract: Tuberculosis is a global health problem. The Mycobacterium bovis Bacille Calmette Guerin (BCG) vaccine has variable efficacy (0–80%) so there is a drive to develop novel vaccines. The cytokine, interferon gamma (IFNγ), is an essential component of the protective response to M. tuberculosis (M. tb) infection and is also produced in response to BCG vaccination. Induction of an IFNγ response is used as a biomarker of successful vaccination in the assessment of new tuberculosis (TB) vaccines. The IFNγ ELISPOT assay provides an important tool for TB research. It is used for both the diagnosis of infection (T.Spot assay), and for the evaluation of the immunogenicity of new TB vaccine candidates in human clinical trials, in the non-human primate (NHP) model of TB infection studies. The ELISPOT assay captures IFNγ produced by peripheral blood mononuclear cells (PBMCs) following specific stimulation, onto a membrane so individual cells can be enumerated and the frequency of responding cells determined. Hence spot forming units (SFU) per 10⁶ cells provide the traditional measure for ELISPOT assays. The discriminatory power of SFU is limited. In some situations, the number of SFU in BCG vaccinated, and unvaccinated, subjects was found to be similar, although the spots were observed to be larger in vaccinated subjects. Spot size potentially provides a measure of the quantity of cytokine produced by individual cells. The AID ELISPOT plate reader software used to determine frequency of spots also has the capability to determine the size of each spot. Consideration of spot size in combination with spot forming units was investigated in our studies of BCG immunogenicity. This additional readout was found to enhance the discriminatory power of the ELISPOT assay, and provide more information on the immune response to BCG vaccination and infection with M.tb.

Abstract: The traditional enzyme-linked immunospot (ELISpot) assay is the gold standard for the enumeration of antigen-specific B cells. Since B cell availability from biological samples is often limited, either because of sample size/volume or the need of performing multiple analyses on the same sample, the implementation of ELISpot assay formats that allow the simultaneous detection of multiple antibody types is desirable. While dual-color ELISpot assays have been described, technical complexities have so far prevented their wide utilization as well as further expansion of their multicolor capability. An attractive solution is to replace the chromogenic reaction of the traditional ELISpot assay with a fluorescent detection system (fluorospot assay). Fluorospot assays using fluorophore-conjugated secondary antibodies in conjunction with fluorescence enhancers, FITC/anti-FITC and biotin/avidin amplification systems and dedicated equipment for spot detection have been developed to enumerate T-cells secreting two or three specific cytokines and, more recently, IgG and IgA antibody-secreting cells (ASCs). We hereby report a method for a multiplex B cell fluorospot assay that utilizes quantum-dot nanocrystals as reporters without further amplification systems or need of dedicated equipment. With this method we simultaneously enumerated HIV-1 gp41 envelope glycoprotein-specific IgG and IgM antibody-secreting cells with sensitivity comparable to that of the traditional ELISpot assay.