Improved cognitive and memory abilities in a patient with Alzheimer's disease treated with activated immune cells: Immune cell therapy may benefit more AD patients.

So far, the pathogenesis of Alzheimer's disease (AD) has not been clarified, nor has patient therapy been satisfactory. Although inheritance dominates the less frequent early-onset AD in young and middle-aged individuals, environmental and immunogenetic factors have been identified in the most frequently occurring late-onset AD of higher-aged individuals, comprising 90% of AD patients. Thorough investigations have detected a prevalence of certain microbes which are known to affect brain activities in the brains of AD patients. This microbial prevalence suggests failing immune responses by immune gene variants against specific microbes. In fact, some immune gene variants have been detected significantly more often in AD patients. Failing immune responses can be corrected by activating immune cells outside the body ("in vitro") for the subsequent therapeutical injections. Activated immune cells digest and present microbial peptides better and differentiate naïve/resting immune cells to powerful effector cells, which can be used for therapy. The patient's activated immune cells can pass the blood-brain barrier and overcome chronic infections in the brain. Furthermore, activated immune cells can secrete a series of neurotrophins for the restoration of neuronal circuits. Based on the encouraging results of immunotherapy in a patient with late-onset AD, we hypothesize that therapy with the patient's activated immune cells would safely benefit many AD patients.

Activated monocytes prime naïve T cells against autologous cancer: vigorous cancer destruction in vitro and in vivo.

It has not been considered so far that antigen-presenting cells (APC) may phagocytose immunogenic material from autologous cancer cells. Assuming the presence of cancer-immunogenic material in APC, we developed a novel autologous priming method that does not require tumour cells or identified peptides. Cancer-immunogenic information came from CD14+ monocytes. When stimulated with CD3-activated T cells, monocytes primed CD3+ CD4+ and CD3+ CD8+ resting/naïve T cells to become powerful effector cells within 24 h. During priming, depletion of CD14+ monocytes but not CD1a+ CD83+ dendritic cells prevented T cell priming. During cancer cell destruction, dendritic cells, but not monocytes, enhanced cancer cell lysis. The cascade-primed (CAPRI) immune cell quartet comprising monocytes, dendritic cells, CD4+ T and CD8+ T cells induced a significant decrease in the number of suppressive CD25(high) FoxP3+ CD4+ T cells. CAPRI cells induced a marked upregulation of MHC class I and class II expression in cancer cells, which is crucial for autoimmune-like lysis. We show in vivo evidence of the CAPRI cell concept in nude mice. In humans, we present circumstantial clinical evidence showing the efficacy of CAPRI cells in an adjuvant treatment attempt for breast cancer patients with metastasis (N = 42). Compared to patients at the Munich Tumor Center (no CAPRI treatment N = 428), almost double the expected number of patients survived 5 years (P =1.36 × 10⁻¹4). The CAPRI method is a safe procedure without negative side effects. High numbers of cancer-specific CAPRI cells can be obtained within a week against different cancer types for efficient adoptive cell therapy.

Associations between Chlamydophila infections, schizophrenia and risk of HLA-A10.

Several microbes have been suspected as pathogenetic factors in schizophrenia. We have previously observed increased frequencies of chlamydial infections and of human lymphocyte antigen (HLA)-A10 in independent studies of schizophrenia. Our aim here was to analyze frequencies of three types of Chlamydiaceae in schizophrenic patients (n=72), random controls (n=225) and hospital-patient controls (n=36), together with HLA-A genotypes. Patients were diagnosed with schizophrenia according to Diagnostic and Statistical Manual of Mental Disorders-IV. Blood samples were collected at the beginning of hospitalization and analyzed with Chlamydiaceae species-specific polymerase chain reaction (PCR). Control panels consisted of randomly selected volunteers and hospitalized, non-schizophrenic patients. We found chlamydial infection in 40.3% of the schizophrenic patients compared to 6.7% in the controls. The association of schizophrenia with Chlamydiaceae infections was highly significant (P=1.39 x 10(-10), odds ratio (OR)=9.43), especially with Chlamydophila psittaci (P=2.81 x 10(-7), OR=24.39). Schizophrenic carriers of the HLA-A10 genotype were clearly most often infected with Chlamydophila, especially C. psittaci (P=8.03 x 10(-5), OR=50.00). Chlamydophila infections represent the highest risk factor yet found to be associated with schizophrenia. This risk is even further enhanced in carriers of the HLA-A10 genotype.

Do dogs harbour risk factors for human breast cancer?

We ask consulting patients regularly whether they keep pets in order to identify zoonotic factors. It became apparent that patients with breast carcinoma (N=69) owned significantly more often dogs but not cats compared to age matched female controls. We compared the frequencies of dog and pet ownership with data from public available statistics on women (N=1320) of the same age group in Bavaria. The most striking result was that more than twice the number of patients kept dogs permanently in the last 10 years and at the time of interrogation as compared to control individuals at the time of interrogation (p=0.0000003, relative risk 3.5). Further internet search on the morbidity of breast carcinoma showed in dogs a protracted course of disease and metastases into lung, liver and bones, resembling the course of disease in human breast cancer. In contrast with this, breast cancer presented in cats a dramatically short course and the main but unusual location of metastasis presents in the hind legs. A recent publication in Norway reported on a high frequency (53.3%) of breast carcinomas in 14,401 investigated dogs. Which transmissible factor or factors come into question? Variants of the mouse mammary tumor virus (MMTV) can productively replicate in human cells and in different animals, including dogs. Many investigators, but not all, could identify MMTV-like sequences in sporadic human breast cancer. MMTV or MMTV-like sequences have not been investigated in canine breast carcinomas until now. It is also conceivable that other microbes from the dog, for example bacteria, could participate in the first steps of carcinogenesis in human. It was recently shown that bartonella species promote vascularization and prevent apoptosis of infected cells with the same methods as helicobacter pylori. Our considerations require further research. Epidemiologic cohort studies and identification of potential carcinogenic microbial factors will prove or disprove our hypothesis that risk factors from dogs could contribute to the carcinogenesis of human breast cancer.

Heat shock protein 70 (Hsp70) stimulates proliferation and cytolytic activity of natural killer cells.

We previously demonstrated that lysis of tumor cells that express Hsp70, the highly stress-inducible member of the HSP70 family, on their plasma membrane is mediated by natural killer (NK) cells. Here, we studied the effects of different proteins of the HSP70 family in combination with interleukin 2 (IL-2) on the proliferation and cytotoxic activity of human NK cells in vitro. Proliferation of NK cells was significantly enhanced by human recombinant Hsp70 (rHsp70) and to a lesser extent by rHsp70homC, the recombinant C-terminal peptide-binding domain derived from Hsp70hom, but not by the constitutive Hsc70 or DnaK, the Escherichia coli analogue of human Hsp70. Even rHsp70 protein alone moderately enhances proliferation and cytolytic activity of NK cells, thus indicating that the stimulatory effect is not strictly dependent on IL-2. NK cells stimulated with rHsp70 protein also exhibit an increased secretion of interferon gamma (IFN-gamma). The phenotypic characterization of NK cells with specificity for Hsp70-expressing tumor cells revealed a CD16dim/CD56bright and increased CD57 and CD94 expression. The cytolytic activity of NK cells also was significantly reduced when a CD94-specific antibody or rHsp70 was added directly before the cytotoxicity assay, whereas other antibodies directed against CD57 and major histocompatibility complex class I molecules or Hsp70 proteins, including Hsc70 and DnaK, did not affect the NK-mediated killing. However, long-term incubation of NK cells with rHsp70 protein enhances not only the proliferative but also the cytolytic response against Hsp70-expressing tumor cells. Our results indicate that the C-terminal domain of Hsp70 protein affects not only the proliferative but also the cytolytic activity of a phenotypically distinct NK cell population with specificity for Hsp70-expressing tumor cells. 1999 International Society for Experimental Hematology.

Epstein-Barr virus-infected B cells of males with the X-linked lymphoproliferative syndrome stimulate and are susceptible to T-cell-mediated lysis.

Primary infection with the Epstein-Barr virus (EBV) results in fatal infectious mononucleosis in up to 70% of males affected by the X-linked lymphoproliferative syndrome (XLP). This rare disease is often associated with diverse natural killer (NK)-, B- and T-cell deficiencies. We describe experiments testing whether the B lymphocytes of affected males play a role in the pathogenesis of XLP due to a low susceptibility to T-cell-mediated immunity. Using reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemistry we detected in these B cells the expression of viral proteins EBNA-1, EBNA-2, EBNA-3A, EBNA-3C, LMP-1 and LMP-2A, which provide targets for cytotoxic T cells. Major histocompatibility complex (MHC) class I, MHC class II and the B7 costimulatory molecule were present on the cell surface. Accordingly, the EBV-infected B cells were lysed in 51Cr-release assays by T lymphocytes sharing MHC determinants with the targets. This MHC-restricted and specific lysis was confirmed in competition experiments using MHC-specific monoclonal antibodies (MAbs) and synthetic peptides. XLP-derived LCLs could also induce MHC class I-restricted memory and cytotoxic T lymphocytes. Thus, these XLP-derived B cells resembled normal LCIs in vitro with respect to induction of EBV-specific cytotoxic T cells (CTL), the ability to present EB viral antigens and the susceptibility to EBV-specific and MHC-restricted CTL-mediated killing. The failure of the immune system to eliminate these virus-infected B cells in XLP is clearly not caused by a B-cell-specific defect.