Advances in Clinical and Experimental Medicine

Title abbreviation: Adv Clin Exp Med
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ISSN 1899–5276 (print)
ISSN 2451-2680 (online)
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Advances in Clinical and Experimental Medicine

2012, vol. 21, nr 3, May-June, p. 289–299

Publication type: original article

Language: English

The Bactericidal Activity of Normal Human Serum Against Enterobacteriaceae Rods with Lipopolysaccharides Possessing O-Antigens Composed of Mannan

Bakteriobójcza aktywność normalnej surowicy ludzkiej wobec szczepów z rodziny Enterobacteriaceae z lipopolisacharydami zawierającymi O-antygeny typu mannanowego

Dorota Tichaczek-Goska1,, Danuta Witkowska2,, Agnieszka Cisowska1,, Stanisław Jankowski1,, Andrzej B. Hendrich1,

1 Department of Biology and Medical Parasitology, Wroclaw Medical University, Wroclaw, Poland

2 Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Wroclaw, Poland

Abstract

Background. The susceptibility of bacteria to the bactericidal activity of serum depends on the structure and organization of the bacterial outer membrane. It is known that the structure of the O-specific polysaccharide chain of lipopolysaccharide (LPS) plays an important role in the resistance of bacteria to host immune defenses.
Objectives. The susceptibility of rods belonging to Enterobacteriaceae family to the bactericidal activity of the normal human serum (NHS) was examined. The mechanisms of complement activation were also investigated.
Material and Methods. The study was carried out on 15 strains containing LPSs with O-specific polysaccharides composed of mannan, belonging to the following species: Citrobacter freundii, C. werkmanii, C. braakii, C. youngae, Hafnia alvei, Escherichia coli and Klebsiella pneumoniae. The levels of C3 and C4 complement components, IgG and IgM immunoglobulin in NHS were examined using specific antibodies. The bactericidal activity of NHS and its preparations (HS50/20, HSMgEGTA) was determined. LPSs from E. coli O8 strains were analyzed by polyacrylamide gel electrophoresis (PAGE) in the presence of sodium dodecyl sulphate (SDS).
Results. Eleven strains were sensitive to NHS bactericidal activity, and four were resistant. Only one group of strains was fully susceptible to NHS action. In three other groups, both sensitive and resistant strains were found. The majority of the strains remained susceptible to NHS activity irrespective of which pathway of serum activity was blocked. All E. coli O8 strains contained smooth-type LPSs.
Conclusion. Strains belonging to the same serotype showed variable susceptibility to the bactericidal action of normal human serum. Two mechanisms of the bactericidal activity of NHS have been identified.

Streszczenie

Wprowadzenie. Podatność bakterii na bakteriobójcze działanie surowicy zależy od struktury i organizacji błony zewnętrznej drobnoustroju. Wiadomo, że struktura polisacharydowego łańcucha O-swoistego odgrywa istotną rolę w oporności bakterii na działanie układu odpornościowego organizmu gospodarza.
Cel pracy. Zbadano podatność pałeczek należących do rodziny Enterobacteriaceae na bakteriobójcze działanie normalnej surowicy ludzkiej (NSL). Określono również mechanizm aktywacji układu dopełniacza oraz jego rolę w procesie lizy komórek przez NSL.
Materiał i metody. Badaniami objęto grupę 15 szczepów mających O-antygeny typu mannanowego, należących do gatunków: Citrobacter freundii, C. werkmanii, C. braakii, C. youngae, Hafnia alvei, Escherichia coli i Klebsiella pneumoniae. Oznaczano stężenia składowych dopełniacza C3 i C4 oraz przeciwciał klasy IgG i IgM w NSL. Określano bakteriobójcze działanie NSL oraz jej preparatów (SL50/20, SLMgEGTA) w stosunku do testowanych pałeczek. Wyizolowano i charakteryzowano elektroforetycznie (SDS-PAGE) lipopolisacharydy (LPS) szczepów E. coli O8.
Wyniki. Jedenaście szczepów bakterii było podatnych, a cztery oporne na działanie NSL. Tylko jedna grupa pałeczek była wrażliwa na działanie NSL. W pozostałych grupach drobnoustrojów, mimo że struktury O-antygenowe były identyczne, odnotowano zarówno podatne, jak i niewrażliwe na NSL szczepy. Większość bakterii nie zmieniła swojego wzoru podatności na surowicę, niezależnie od tego, którą z dróg układu dopełniacza inaktywowano. Wszystkie szczepy E. coli O8 miały LPS typu gładkiego.
Wnioski. Szczepy bakteryjne należące do jednego gatunku i serotypu wykazują zróżnicowanie w podatności na lityczne działanie białek układu dopełniacza. Odnotowano dwa mechanizmy bakteriobójczego działania NSL wobec testowanych pałeczek.

Key words

mannan, O-antigen, human serum, bactericidal effect, complement system

Słowa kluczowe

mannan, O-antygen, ludzka surowica, działanie bakteriobójcze, układ dopełniacza

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