Laboratory for Evolutionary and Ecological Immunology

Doctoral theses

Hana Velová (Bainová) (2020): Intraspecific and interspecific variability in bacterial detection mediated by Toll-like receptors in birds

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Barbora Bílková (2018): Variability of chicken breeds in egg antimicrobial peptides

Master theses

Daniel Divín (2020): Inflammation-associated changes in cannabinoid receptor expression in avian brain

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Sylvie Dlugošová (2020): Variability in selected haematological traits related to gastrointestinal microbiota in parrots (Psittaciformes)

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Tereza Krajzingrová (2019): Effect of ageing on haematological and immunological traits in great tit (Parus major)

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Lucie Buchtová (2016): Relatedness and gene flow between breeds of the domestic fowl

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Martin Těšický (2016): Trans-species polymorphism in selected genes of innate immunity in tits (Paridae)

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Anežka Pojezdná (2015): MHC class I and MHC class II allele variability in domestic fowl breeds

Barbora Bílková (2014): Variability in health state of mice in Mus musculus musculus and Mus musculus domesticus hybrid zone

Zuzana Bainová (2013): Polymorphism of TLR2/TLR1 and TLR2/TLR6 heterodimers in wild-derived house mouse inbred strains

Contrary to the classical mouse inbred strains with unnatural genetic variability, wild- derived strains offer a more suitable model for evolutionary immunology. Toll-like receptors (TLRs) belong to initial detectors of invading pathogens. Although TLRs recognise conserved structures they were shown to be polymorphic. This polymorphism is associated with various diseases. In my thesis, I describe variability of Tlr1, 2 and 6 in 24 inbred strains derived from two subspecies of house mouse (Mus m. musculus and M. m. domesticus). These Tlrs exhibit different levels in variability among the strains. In Tlr1 the polymorphic sites are spread along the whole exodomain. Tlr6 is quite conserved (a lower amount of substitutions located far from the binding region and with minor modifications in the amino acid residue properties). Tlr2, on the contrary, contains some substitutions with substantial alternations of residue properties that are located within or nearby the binding region and the subspecies differ at these sites. All alleles of M. m. domesticus and M. m. musculus, except for Tlr1 PWD, Tlr2 STAIL, are phylogenetically separated. The strains and the subspecies vary in the production of IL-1β, IL-12 a NO after stimulation by TLR1, 2 and 6 ligands. This trend is, however, presumably influenced by the effect of particular lines (e.g. BULS in the case of IL-1β). The results of my thesis imply independent co-evolution of TLR2 with pathogens in these subspecies.

Hana Bainová (2011): The Influence of Toll-like Receptor 4 Polymorphism on Condition and Ornamentation in Great Tits

Host-parasite co-evolution belongs to the most important evolutionary relationships that shape natural and sexual selection. Parasites pose permanent selective pressure on their hosts. Toll-like receptors (TLRs) as a part of innate immunity are involved in mechanisms of a first immunological barrier which has to be overcome by parasites. These receptors play a key role in primary detection of pathogen-associated molecular patterns (PAMPs) and, hence, are responsible for early triggering of effector immunological mechanisms and for co-activating adaptive immunity. Several studies revealed that TLR4 may represent a suitable model molecule for host-parasite co-evolution studies. TLR4 interacts directly with several PAMPs and structural variability in this receptor was shown to affect host resistance to various diseases. Thus, there is potential for occurrence of parasite-mediated natural and sexual selection. Contrary to the number of fish and mammalian TLRs described, avian inter- and intraspecific TLR variability is only very insufficiently explored. This is especially true for passerine birds. In my diploma thesis I therefore provide the first description of the complete Tlr4 translated region in a non-model free-living bird, great tit (Parus major), predict structure of the protein product of this gene and analyse its population polymorphism. To assess evolutionary forces acting on the great tit TLR4 I described the TLR4 also in several other passerine species. These data were used to investigate influence of the TLR4 polymorphism on condition-related traits in great tit. I found that that one particular amino acid substitution, Q549R, is associated with expression of plumage ornamentation. In both sexes individuals bearing this substitution express narrower melanin-pigmented black breast stripes and lighter carotenoid-pigmented yellow breast colouration. To my knowledge, this is the first evidence for possible association between TLR polymorphism and ornamental colouration in animals. In general, our data show influence of innate immunity on ornamental signalling and support indicator model of sexual selection.

Bachelor theses

Klára Hančová (2019): Evolution of immune response driven by increasing pathogen virulence

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Daniel Divín (2018): Evolution of host specificity in avian influenza

Sylvie Dlugošová (2017): Evolutionary dynamics of the CD45 molecule

Matěj Pavlík (2016): Changes in expression of immunological signalling molecules during senescence

Lucie Buchtová (2014): Usage of molecular genetics in investigation of origin of domesticated animals

Martin Těšický (2014): Trans-species polymorphism in immune genes at wildlife animals

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Julie Vacková (2014): Effect of parasites on haematological traits in vertebrates

Anežka Pojezdná (2013): Changes in expression of immune-related genes after induction of inflammation in birds

Barbora Bílková (2012): Evolution of molecular interaction between host and parasite

Coevolution between host and parasite is a long-term object of scientific interest, mostly because of negative influence of parasites and human need to defend against it., The evidence concerning host-parasite coevolution mechanisms is, however, still incomplete and various models and theories are considered separately, out of the general concept. Beside generally well-known theories such as the “Red queen” theory or the coevolution “arms race” theory, several other models were proposed, e.g. the “gene for gene” model or the “matching allele” model, which describe detailed aspects of host-parasite coevolution and principles of genetic- variability maintenance in their interaction. Although there are many studies mapping reciprocal evolution of hosts and parasites, the connection between empirical evidence and theoretical models is often missing. Therefore, this thesis covers theory on host-parasite evolutionary interactions and provides examples of host-parasite systems and molecules, which correspond to behavior of described models.

Hana Paclíková (2012): Associations of innate immunity gene polymorphism with inflammatory diseases in mammals

The immune system of vertebrates is very complicated and complex structure, helping individuals in their daily struggle for survival. His innate and adaptive components are involved in a number of mechanisms, which have been optimized in evolution to provide most efficient defense to the organism with the lowest possible costs and consequences. Despite this fact, there are many diseases caused by the malfunction of the immune system. This thesis is focused on these diseases and on non-specific part of the immune system, particularly Toll-like receptors that are responsible for recognizing the signals of potential danger to the organism. These receptors are often highly polymorphic, which may lead to the structural changes influencing the protein function. The variability associated with the pro-inflammatory diseases is discussed with specific examples being provided. Immunopathology and autoimmune diseases are a major problem not only in the human community but also of many farm and free-living animals. Detailed surveys and mapping of polymorphism of innate immunity receptors may be of a great importance to solving zoohygienic and veterinary problems as well as protection of populations of wild mammals.

Jitka Poplová (2011): Application of haematological methods in zoological studies

Zuzana Bainová (2010): Evolutionary implications of innate immunity receptors polymorphism

Hana Bainová (2008): Methodological aspects of cell immunity measurements in evolutionary ecology studies

The capability to defend oneself against parasitic infections is absolutely crucial for survival in the real world. Therefore the considerable part of current evolutionary-ecological studies try to describe and explain differences among individuals and species that are based in capability of mounting an effective immune reaction to pathogenic stimulation. There are many methods of measuring immune reaction based on different principles. Phytohaemagglutinin test (PHA, Phytohaemagglutinin) of skin hypersensitivity is the one which is used most frequently mainly because of its easy application in field. Understanding the immunological mechanisms, on which PHA test is based, is important for appropriate planning of experiments as well as for following correct interpretation of the data. However, the functional nature of induced swelling reaction still remains very unclear. In this work I have resumed the current knowledge on structural and biological features of PHA. I also present aggregate table of its usage both in in vitro and in vivo studies, including analysis of methodological aspects of PHA skin test. Unlike expectation settled for the experimental part of my work indicates that the activation of immune reaction to PHA and following swelling can be caused by other mechanisms than lymphocytes too. We have found that PHA skin hypersenzitivity reaction depends not just on the ability of L subunits to stimulate lymphocyte proliferation. The application of PHA consisting only of isomer E subunits, which is conductive to erytagglutination, evoked similar, but quantitatively considerably significantly stronger reaction in the tested animals than pure L tetramers. It seems that the hypersensitivity reaction to PHA-P might therefore simulate state similar to immune response to injury connected with blood coagulation. Although our results relativize a bit the interpretation value of PHA test, I still belive that these data can be really useful, especially in the primary screening of differences in nonspecific skin hypersensitivity among various individuals.