Klára Kubelková

University of Defence

Author Statistics

Papers

Citation

H-Index

i-10 index

Research Trends

Author Order

Document Type

Co-Authors

Aleš Macela

University of Defence

Jaroslav Pejchal

University of Defence

ZŠ

Zuzana Šinkorová

University of Defence

Aleš Tichý

National Centre of Scientific Research "Demokritos"

Zuzana Kročová

University of Defence

Lenka Zárybnická

University of Defence

Jiří Stulík

University of Defence

Jiřina Vávrová

University of Defence

Kamil Kuča

University Hospital Hradec Králové

Lenka Plzakova

University of Defence

Cooperative Institutions

University of Defence

Charles University

University Hospital Hradec Králové

University of Hradec Králové

University of Pardubice

University of Defence

Czech Academy of Sciences

Jan Evangelista Purkyně Military Medical Academy

Veterinary Research Institute

Military Medical Academy

Author Statistics

Papers

Citation

H-Index

i-10 index

Research Field

Francisella and Antibodies

Microorganisms (2021)

Klára Kubelková Aleš Macela

Immune responses to intracellular pathogens depend largely upon the activation of T helper type 1-dependent mechanisms. The contribution of B cells to establishing protective immunity has long been underestimated. Francisella tularensis, including a number of subspecies, provides a suitable model for the study of immune responses against intracellular bacterial pathogens. We previously demonstrated that Francisella infects B cells and activates B-cell subtypes to produce a number of cytokines and express the activation markers. Recently, we documented the early production of natural antibodies as a consequence of Francisella infection in mice. Here, we summarize current knowledge on the innate and acquired humoral immune responses initiated by Francisella infection and their relationships with the immune defense systems.

Francisella

Intracellular parasite

Tularemia

Humoral immunity

10.3390/microorganisms9102136

Cite

Citations (10)

Early infection-induced natural antibody response

Scientific Reports (2021)

Klára Kubelková Tomáš Hudcovic Hana Kozáková Jaroslav Pejchal Aleš Macela

Abstract There remains to this day a great gap in understanding as to the role of B cells and their products—antibodies and cytokines—in mediating the protective response to Francisella tularensis , a Gram-negative coccobacillus belonging to the group of facultative intracellular bacterial pathogens. We previously have demonstrated that Francisella interacts directly with peritoneal B-1a cells. Here, we demonstrate that, as early as 12 h postinfection, germ-free mice infected with Francisella tularensis produce infection-induced antibody clones reacting with Francisella tularensis proteins having orthologs or analogs in eukaryotic cells. Production of some individual clones was limited in time and was influenced by virulence of the Francisella strain used. The phylogenetically stabilized defense mechanism can utilize these early infection-induced antibodies both to recognize components of the invading pathogens and to eliminate molecular residues of infection-damaged self cells.

Francisella

Tularemia

Intracellular parasite

10.1038/s41598-021-81083-0

Cite

Citations (4)

HOST PATHOGEN INTERACTION FORUM HAS ACHIEVED INTERNATIONAL RECOGNITION

MMSL (2018)

Klára Kubelková

Source

Cite

Citations (0)

FRANCISELLA TULARENSIS - 100 YEARS: TULAREMIA RESEARCH IN FORMER CZECHOSLOVAKIA AND IN THE CZECH REPUBLIC

Military Medical Science Letters (2012)

Aleš Macela Jiří Stulík Zuzana Kročová Michal Kroča Klára Kubelková

The history of national tularemia research started in 1936 when the first outbreak was recognized in south-east Moravia.Since then in average about one hundred cases have been recorded annually.As tularemia was endemic in former Czechoslovakia, three research groups which concentrated on this disease were formed during decades.The first two groups have worked from sixties and were associated with Jiri Libich (Prague) and Darina Gurycova (Bratislava).The third group which concentrated on the research of natural foci started during late seventies in Valtice (Zdenek Hubalek).The experimental research was, and still is, mainly associated with military research, recently with the Proteomic Center (Faculty of Military Health Sciences, University of Defence) in Hradec Kralove.This center opens molecular approaches to the analysis of Francisella tularensis microbes on one side and the studies on mutual host-pathogen interaction on the other side.One of the significant aims of the research is searching for the new typing and diagnostic markers of Francisella tularensis for the military and medical practice.Thus, scientists from former Czechoslovakia and the Czech Republic contributed significantly to current knowledge on Francisella pathogenesis and their results were highly appraised by international scientific community.

Tularemia

Francisella

Zoonotic disease

10.31482/mmsl.2012.006

Cite

Citations (4)

Development of tularemic scFv antibody fragments using phage display

Open Life Sciences (2010)

Klára Kubelková Aleš Macela

Abstract Polyclonal antibodies, as well as monoclonal antibodies are efficacious in providing protective immunity against Francisella tularensis. This study demonstrates the application of phage display libraries for the construction of monoclonal antibodies against F. tularensis. Novel single-chain fragment variable (scFv) antibodies were generated against a whole bacterial lysate of F. tularensis live vaccine strain using the human single fold scFv libraries I (Tomlinson I + J). A total of 20 clones reacted with the bacterial cell lysate. Further, the library contains two clones responsive to recombinant lipoprotein FTT1103Δsignal (F. tularensis subsp. tularensis Schu S4), which was constructed without a signal sequence. These positively-binding scFvs were evaluated by scFv-phage enzyme-linked immunosorbent assay (ELISA). Then, positive scFvs were expressed in a soluble form in Escherichia coli HB2151 and tested for positive scFvs by using scFv-ELISA.

Polyclonal antibodies

Single-chain variable fragment

Phagemid

10.2478/s11535-010-0015-3

Cite

Citations (0)

Modulation of ionizing radiation-induced effects by NU7441, KU55933 and VE821 in peripheral blood lymphocytes

Journal of Applied Biomedicine (2015)

Adéla Kmochová Aleš Tichý Lenka Zárybnická Zuzana Šinkorová Jiřina Vávrová

We evaluated the impact of ataxia-telangiectasia mutated kinase inhibitor KU55933, DNA-dependent protein kinase inhibitor NU7441 and ataxia telangiectasia and rad3-related kinase inhibitor VE821 in human peripheral lymphocytes in vitro. The lymphocytes were divided into 5 groups: non-irradiated control, irradiated group (2 Gy) and 3 groups pretreated with inhibitors 30 min before irradiation. We used flow cytometry to evaluate phosphorylated H2AX (γ-H2AX) and cytotoxicity (Apoptest). Micronucleus assay was used to assess genotoxicity. After irradiation, γ-H2AX, incidence of micronuclei (MN), nucleoplasmatic bridges (NPBs) and nuclear buds in binuclear cells, MN in mononuclear cells and apoptosis were increased. KU55933 decreased γ-H2AX and inhibited ionizing radiation-induced cytotoxicity. NU7441 showed no effect on γ-H2AX but it significantly increased MN and NPBs in binuclear cells and apoptosis. VE821 decreased γ-H2AX, whereas genotoxicity and cytotoxicity were not affected. In conclusion, KU55933 protected lymphocytes, which might be employed to preserve the immune system during anticancer therapy. NU7441 radiosensitized lymphocytes, thus, undesirable side effects toward immune system could be expected. VE821 showed decrease of γ-H2AX with no radiosensitizing effects in our model likely due to p53 positive status, which underlies the concept of its application in p53 negative environment.

10.1016/j.jab.2015.07.001

Cite

Citations (12)

B cell subsets are activated and produce cytokines during early phases of Francisella tularensis LVS infection

Microbial Pathogenesis (2014)

Lenka Plzakova Klára Kubelková Zuzana Kročová Lenka Zárybnická Zuzana Šinkorová

Tularemia

CD80

CD86

10.1016/j.micpath.2014.08.009

Cite

Citations (16)

Možnosti purifikace tularemického proteinu IglC a jeho vazebných partnerů

Klára Kubelková

Source

Cite

Citations (0)

Entry of Francisella tularensis into Murine B Cells: The Role of B Cell Receptors and Complement Receptors

PLoS ONE (2015)

Lenka Plzakova Zuzana Kročová Klára Kubelková Aleš Macela

Francisella tularensis, the etiological agent of tularemia, is an intracellular pathogen that dominantly infects and proliferates inside phagocytic cells but can be seen also in non-phagocytic cells, including B cells. Although protective immunity is known to be almost exclusively associated with the type 1 pathway of cellular immunity, a significant role of B cells in immune responses already has been demonstrated. Whether their role is associated with antibody-dependent or antibody-independent B cell functions is not yet fully understood. The character of early events during B cell–pathogen interaction may determine the type of B cell response regulating the induction of adaptive immunity. We used fluorescence microscopy and flow cytometry to identify the basic requirements for the entry of F. tularensis into B cells within in vivo and in vitro infection models. Here, we present data showing that Francisella tularensis subsp. holarctica strain LVS significantly infects individual subsets of murine peritoneal B cells early after infection. Depending on a given B cell subset, uptake of Francisella into B cells is mediated by B cell receptors (BCRs) with or without complement receptor CR1/2. However, F. tularensis strain FSC200 ΔiglC and ΔftdsbA deletion mutants are defective in the ability to enter B cells. Once internalized into B cells, F. tularensis LVS intracellular trafficking occurs along the endosomal pathway, albeit without significant multiplication. The results strongly suggest that BCRs alone within the B-1a subset can ensure the internalization process while the BCRs on B-1b and B-2 cells need co-signaling from the co receptor containing CR1/2 to initiate F. tularensis engulfment. In this case, fluidity of the surface cell membrane is a prerequisite for the bacteria's internalization. The results substantially underline the functional heterogeneity of B cell subsets in relation to F. tularensis.

Tularemia

B-1 cell

Internalization

Francisella

10.1371/journal.pone.0132571

Cite

Citations (16)

Nové možnosti pasivní imunoprofylaxe intracelulárních bakteriálních infekcí

MMSL (2008)

Klára Kubelková Aleš Macela

Infekce způsobene intracelularnimi bakterialnimi patogeny jsou považovany obecně za važný problem veřejneho zdravotnictvi. Vzhledem k neexistenci vakcin pro humanni použiti neexistuje u větsiny infekci ani žadna vakcinacni strategie. Informace ziskane z nových studii nabizeji možnost využiti polyklonalnich nebo monoklonalnich protilatek samotných ci v kombinaci s cytokiny k pasivni imunoprofylaxi těchto zavažných infekcnich onemocněni.

Source

Cite

Citations (0)