Hansjörg Thude

Allele frequencies of human platelet antigen 1, 2, 3, and 5 systems in patients with chronic refractory autoimmune thrombocytopenia and in normal persons.

Background and Objectives: Chronic refractory autoimmune thrombocytopenic purpura (AITP) is an autoimmune disorder due to autoantibodies against platelet glycoproteins (GP). Human platelet alloantigenic (HPA) systems are distributed to different platelet GPs. We carried out genotyping of diallelic HPA–1, –2, –3, and –5 systems to clarify potential associations between HPA alleles and the development of chronic refractory AITP. Patients and Methods: DNA was isolated from 33 unrelated German patients with chronic refractory AITP and from 80 randomly selected German blood donors to determine the phenotype and allele frequencies for the HPA–1, –2, –3, and –5 systems. Fragments carrying the polymorphic sequences corresponding to those alleles were amplified by the polymerase chain reaction and further characterized by restriction analysis. Results: Whereas HPA–1, –3, and –5 allele frequencies were identical in 33 patients with chronic refractory AITP and in controls, HPA–2 allele frequencies showed a statistically significant difference (p = 0.017). In our group of patients, the HPA–2a allele frequency was 100%, but HPA–2b was not seen. In contrast, the allele frequency of HPA–2a in the control group was 92% (n = 147), and in HPA–2b it was 8% (n = 13). Conclusion: This study suggests an association between the HPA–2a allele and chronic refractory AITP. The HPA–2a allele may be involved in the formation of an AITP–specific autoepitope.

De novo mTOR inhibitor-based immunosuppression in ABO-incompatible kidney transplantation

ABO‐incompatible (ABOi) kidney transplantation (KTx) has become an accepted therapeutic option in renal replacement therapy for patients without a blood group‐compatible living donor. Using different desensitization strategies, most centers apply B‐cell depletion with rituximab and maintenance immunosuppression (IS) with tacrolimus and mycophenolic acid. This high load of total IS leads to an increased rate of surgical complications and virus infections in ABOi patients. Our aim was to establish ABOi KTx using an immunosuppressive regimen, which is effective in preventing acute rejection without increasing the risk for viral infections. Therefore, we selected a de novo immunosuppressive protocol with low‐dose calcineurin inhibitor and the mTOR inhibitor everolimus for our ABOi program. Here, we report the first 25 patients with a complete three‐yr follow‐up treated with this regimen. Three‐yr patient survival and graft survival were 96% and 83%. The rate of acute T‐cell‐mediated rejections was low (12%). Cytomegalovirus (CMV) infection was evident in one patient only (4%). Surgical complications were common (40%), but mild in 80% of cases. We demonstrate that ABOi KTx with a de novo mTOR inhibitor‐based regimen is feasible without severe surgical or immunological complications and a low rate of viral infections.

Treatment of a Patient with Chronic Immune Thrombocytopenic Purpura with Rituximab and Monitoring by Flow Cytometric Analysis

Treatment modalities of patients with chronic immune thrombocytopenic purpura (ITP) include the administration of intravenous immunoglobulins (IVIG), corticosteroids, anti-D(Rh) immunoglobulin (anti-D), and splenectomy. Approximately 25–30% of patients with chronic ITP do not respond to established therapeutic regimens. We describe a 19-year-old patient with chronic ITP refractory to standard therapies treated with rituximab (anti-CD20 antibody). Initially, the therapy with rituximab appeared to be successful; however, the patient relapsed after a surveillance of 57 weeks documenting that the rituximab therapy has failed. Flow cytometric analyses during and after the administration of rituximab revealed new aspects of monitoring rituximab therapy.

Association between a gain-of-function variant of PTPN22 and rejection in liver transplantation.

Background The protein tyrosine phosphatase nonreceptor 22 gene (PTPN22) encodes a strong T-cell regulator called lymphoid protein tyrosine phosphatase. Previously, PTPN22 was described as a susceptibility gene for autoimmunity because it contains single nucleotide polymorphisms (SNPs) associated with several autoimmune diseases. One SNP (rs2476601; 1858G>A) has emerged as a particularly potent risk factor for autoimmunity. We address the question whether PTPN22 polymorphisms are also associated with acute rejection after liver transplantation. Methods We investigated the influence of six PTPN22 SNPs on the susceptibility to acute liver allograft rejection. Consequently, we carried out a retrospective study genotyping 345 German liver recipients at six SNP loci, which include rs2488457 (−1123G>C), rs33996649 (788C>T), rs2476601 (1858G>A), rs1310182 (−852A>G), rs1217388 (−2200G>A), rs3789604 (64434T>G). Our study enrolled 165 recipients who did not develop rejection, 123 who showed one rejection episode, and 57 patients who suffered from multiple acute rejections after transplantation. Results The 1858A allele containing genotypes (GA+AA) and the 1858A allele had a significantly higher frequency in the group of patients with multiple rejection episodes (35.1% and 18.4%) compared to rejection-free patients (15.8% and 7.9%; P=0.022 and 0.023). In contrast, we could not detect any association between rejection and the other tested SNPs. Additionally, we identified one haplotype contributing to risk of multiple rejections, however, exhibiting no stronger impact than the 1858A allele alone. Conclusion We conclude that the 1858G>A SNP may confer susceptibility to multiple acute liver transplant rejections in the German population.

Lack of association between CTLA-4 and PDCD1 polymorphisms and acute rejection in German liver transplant recipients.

Cytotoxic T-lymphocyte-associated antigen 4 (CTLA-4) and programmed cell-death 1 (PDCD1) are two genes encoding coinhibitory immunoreceptors that are involved in regulation transplant rejection and tolerance induction. Thus, CTLA-4 and PDCD1 may be good candidate genes to evaluate in liver transplant rejection. In this retrospective study, we investigated whether four functional single nucleotide polymorphisms (SNP) of the CTLA-4 gene and PDCD1 gene were associated with susceptibility to liver transplant rejection. The SNPs -1772T>C (rs733618), -1661A>G (rs4553808) of the CTLA-4 gene, and the SNPs 7146G>A (rs11568821), 7209C>T (rs41386349) of the PDCD1 gene were genotyped by polymerase chain reaction allele specific restriction enzyme analysis (PCR-ASRA) in 100 liver recipients with acute rejection, 104 liver transplant recipients without acute rejection and 100 healthy control individuals. For the selected SNPs we did not detect any significant difference in genotypic and allelic frequencies between liver transplant recipients with and without acute rejection. In conclusion, our results suggest that the tested SNPs may not be associated with susceptibility to acute liver transplant rejection in a Caucasian population.

CD34+ and CD133+ Primitive Stem Cell Expression in Peripheral Blood: Considering Gender, Age, and Smoking

Background: The number of primitive progenitor cells (pPC) in healthy individuals, in correlation to age, gender, and smoking status, has not yet been thoroughly elucidated. Material and Methods: The pPC from a collective of 168 healthy blood donors aged 18–61 years was investigated using flow cytometric analysis. In addition, the pPC of 20 subjects were studied once a month for half a year to determine the extent of physiological variation of pPC within a single individual. Results: We demonstrated a statistically significant difference (p = 0.005) in the numbers of pPC in men (836,100/l) versus women (583,850/l). No statistical difference was found between younger and older donors or between smokers and non-smokers, both overall and within a single gender. The extent of physiological variation in pPC was lower than 20% in 2 individuals, 18 individuals exhibited amplitudes greater than 20%. Conclusion: We conclude that the number of pPC in healthy individuals was primarily determined by gender as an operative factor. It seems that age and smoking status are of minor importance. Furthermore, our data demonstrate strong variability in the expression of pPC within a single individual. This may be influenced by varying physiological and environmental factors.

Book Review · Buchbesprechung

‘Dirty Medicine. The Handbook’, the latest book from the British investigative writer Martin J. Walker, does a good job of summing itself up in its long subtitle: ‘Dirty Medicine. The Handbook. Of Doctors, Epidemiologists, Researchers, Advisers, Insurance Scammers, Quackbusters, Bad Journalists, Corporate Scientists, Industry Shills, and Other Agents Who Do Now, or Have in The Past Worked For, Sided With, Supported or Simply Been a Part of Corporate Interests, Manufacturing, Pushing, or Promoting, Products or Ideologies Antagonistic to The Private and Public Health of Workers, Consumers, Citizens, Patients, Alternative Practitioners and Science in The Public Interest; Who They Are, How to Recognise Them and Their Organisations, and What to Do About Them.’ The handbook, an extension and reworking of his previous book ‘Dirty Medicine. Science, Big Business and the Assault on Natural Health Care’, provides a who’s who of individuals, groups and websites with a history of attacking complementary and alternative medicine (CAM), particularly in Britain. Starting with an overview, he links the rise of the ‘health fraud’ movement, ‘quackbusters’ and ‘skeptics’ with the rise of corporate science, noting that science is no longer the domain only of scientists, but of public relation companies who provide ‘trained mercenaries to fight the sciences battles’. Continuing with the language of war, he describes the battle tactics of the pharmaceutical industry as akin to those initiated and perfected by the tobacco and asbestos industries. Writing of an organised plan against natural medicine, he states that, as there is no major profit in natural medicine, it has to be ‘controlled and destabilised’ to protect pharmaceutical profits, which increased when medical science became more focused on problems to be fixed rather than the patient as a person. In alphabetical order he gives background information ranging from a line to a page on what he calls ‘health corporatists’: individuals, organisations, ad hoc groups and websites. Pharmaceutical company, personal, and committee links, and their history in relation to attacks against individuals or groups involved in CAM, are outlined. Following these chapters he provides a counterbalance: the antidote groups and individuals historically and currently battling for freedom of treatment and exposing the tactics behind CAM censorship. Some have had their careers damaged or destroyed in the process. In the closing chapter, entitled ‘Organising and Fighting Back’ he pulls no punches in relation to how serious he considers the war against CAM, warning against underestimating the dangers and the lengths to which pharmaceutical companies are willing to go to protect their profits, stating ‘we can define our enemy very simply as those who consistently use lies and manipulation to put profits before people and corporations before communities’. I suspect there are those who will consider much of Walker’s writing as the ravings of a conspiracy theorist who sees apparent links everywhere and who demonises the pharmaceutical industry. In the current climate, however, his work is not so easy to dismiss. Welcoming genuine and honest scientific debate within the scientific community, his major objections are to the often undisclosed conflicts of interest which exist in the media, industry, academia and government, and the way in which the damage caused by allopathic medicine is swept under the carpet. A recent example of this was the failure of BMJ editor Fiona Godlee to disclose the journal’s financial relationship with Merck and GlaxoSmithKline (both measles, mumps and rubella (MMR) vaccine manufacturers) in relation to her editorial and commissioning of 3 articles by a journalist about the Andrew Wakefield case. Wakefield, an English research doctor, lost his licence to practice following a suggestion (in a co-authored paper) that further investigations were needed to see if there was a link between the MMR vaccine and gastrointestinal disease/developmental regression [1]. In her response to criticism, Godlee stated ‘We didn’t declare these competing interests because it didn’t occur to us to do so’ [2]. Her subsequent declaration of advertising and sponsorship income from Merck and GlaxoSmithKline still failed to disclose the partnership agreement between the BMJ and Merck providing online education. While it is within the realms of possibility that the failure of the BMJ to disclose their links to companies that benefitted from Wakefield being considered guilty was an innocent (albeit sloppy) mistake by a journal editor, it is this lack of transparency that, unfortunately, creates a need for research such as that found in the handbook. Anyone who has been confused by the constant rejection of a good research paper, who has been refused the opportunity to rebut criticism in the letters page, or whose comments in a public forum were deleted might better understand the reasons by following the money or political links, some of which are outlined there. Although every link to pharmaceutical companies cannot be automatically considered manipulative and some critics of CAM are driven by an