Scientists from the German Cancer Research Center and the National Center for Tumor Diseases (NCT) Heidelberg have developed a sequencing method with which they can precisely analyze the human T cell receptor repertoire. This makes it possible to study how the immune system responds to disease.
Every human being is unique – especially in molecular terms: every 100th to 300th building block in the genome differs from one human to another. Molecular individuality is particularly pronounced at the level of the genetic material for the immune system’s receptor proteins.
Among these very diverse receptors are T cell receptors as well as antibodies. “This immune receptor diversity is immensely important because it allows exact recognition of the largest possible number of different exogenous substances,” explains Manfred Schmidt from the National Center for Tumor Diseases (NCT) Heidelberg.
The extreme diversity of the T cell receptors has a genetic basis. Each T cell, however, has receptors of a single specificity only. “The specificity and the frequency of the receptor genes together provide an exact picture of the individual’s T cell repertoire. From the frequency with which individual T cell clones occur we can draw conclusions about diseases and the success of therapy,” says Schmidt.
Analyzing the highly variable segments of the T cell receptor genes and accurately determining their frequency is a major scientific challenge. The scientists in the groups of Manfred Schmidt and Christof von Kalle, Director of the NCT, have succeeded in developing a new method* with which they can provide precise sequence analyses of the T cell repertoire.
The two initial authors, Eliana Ruggiero and Jan Nicolay, first tested the method in immune cells from the blood of six healthy donors. As expected, the samples they studied displayed polyclonality: the scientists found thousands of different T cell clones in each of the donors.
By studying the blood of persons infected with the widespread cytomegalovirus (CMV), they were able to elucidate how an infection affects the T cell repertoire (TCR). The researchers treated these blood cells with a mixture of peptides representing the virus’s various antigens, thus simulating the effect of a vaccine. Shortly after the peptides were added, the proportion of those T cells directed against CMV increased, reaching almost 100 percent within about two weeks.
On the basis of TCR investigations in the blood of ten patients suffering from an aggressive T cell lymphoma, the researchers were able to measure by how much the disease reduced the normal T cell repertoire.
“With the new amplification and sequencing method, we can now see, for instance, how the immune system of patients with a serious infection combats the pathogens,” explains Christof von Kalle. “The same approach can be taken in patients with autoimmune diseases to determine which T cell clones attack endogenous tissues; doctors can observe the status of the malignancy in lymphoma patients.” For cancer physicians such as von Kalle, the use of this method in tumor immune therapy is particularly promising: they can now determine the DNA sequence of the T cell receptors that are most effective against the tumor and possibly even artificially replicate them. The huge advances made in the sequencing of DNA and RNA – the authors agree – will in future help medical science gain an even deeper understanding of the human immune system.
* TCR ligation anchored-magnetically captured PCR (TCR-LA-MC PCR)
Eliana Ruggiero, Jan P. Nicolay, Raffaele Fronza, Anne Arens, Anna Paruzynski, Ali Nowrouzi, Gökçe Ürenden, Christina Lulay, Sven Schneider, Sergij Goerdt, Hanno Glimm, Peter H. Krammer, Manfred Schmidt, Christof von Kalle: High-resolution analysis of the human T-cell receptor repertoire. Nature Communications 2015, DOI: 10.1038/ncomms9081
The German Cancer Research Center (Deutsches Krebsforschungszentrum, DKFZ) with its more than 3,000 employees is the largest biomedical research institute in Germany. At DKFZ, more than 1,000 scientists investigate how cancer develops, identify cancer risk factors and endeavor to find new strategies to prevent people from getting cancer. They develop novel approaches to make tumor diagnosis more precise and treatment of cancer patients more successful. The staff of the Cancer Information Service (KID) offers information about the widespread disease of cancer for patients, their families, and the general public. Jointly with Heidelberg University Hospital, DKFZ has established the National Center for Tumor Diseases (NCT) Heidelberg, where promising approaches from cancer research are translated into the clinic. In the German Consortium for Translational Cancer Research (DKTK), one of six German Centers for Health Research, DKFZ maintains translational centers at seven university partnering sites. Combining excellent university hospitals with high-profile research at a Helmholtz Center is an important contribution to improving the chances of cancer patients. DKFZ is a member of the Helmholtz Association of National Research Centers, with ninety percent of its funding coming from the German Federal Ministry of Education and Research and the remaining ten percent from the State of Baden-Württemberg.