Endocrine Microbiology : A transdisciplinary approach of the human physiology

Dominique M., Lucas N., Legrand R., Bouleté I., Bôle-Feysot C., Deroissart C., Léon F., Nobis S., do Rego J., Lambert G., Déchelotte P.

CLPB (Caseinolytic peptidase B) protein is a conformational mimetic of α-MSH, an anorectic hormone. Previous in vivo studies have already shown the potential effect of CLPB protein on food intake and on the production of peptide YY (PYY) by injection of E. coli wild type (WT) or E. coli ΔClpB. However, until now, no study has shown its direct effect on food intake. Furthermore, this protein can fragment naturally. Therefore, the aim of this study was (i) to evaluate the in vitro effects of CLPB fragments on PYY production; and (ii) to test the in vivo effects of a CLPB fragment sharing molecular mimicry with α-MSH (CLPB25) compared to natural fragments of the CLPB protein (CLPB96). To do that, a primary culture of intestinal mucosal cells from male Sprague–Dawley rats was incubated with proteins extracted from E. coli WT and ΔCLPB after fragmentation with trypsin or after a heat treatment of the CLPB protein. PYY secretion was measured by ELISA. CLPB fragments were analyzed by Western Blot using anti-α-MSH antibodies. In vivo effects of the CLPB protein on food intake were evaluated by intraperitoneal injections in male C57Bl/6 and ob/ob mice using the BioDAQ® system. The natural CLPB96 fragmentation increased PYY production in vitro and significantly decreased cumulative food intake from 2 h in C57Bl/6 and ob/ob mice on the contrary to CLPB25. Therefore, the anorexigenic effect of CLPB is likely the consequence of enhanced PYY secretion.

Perraud Q., Kuhn L., Fritsch S., Graulier G., Gasser V., Normant V., Hammann P., Schalk I.J.

Iron is an essential nutrient for bacterial growth and the cause of a fierce battle between the pathogen and host during infection. Bacteria have developed several strategies to access iron from the host, the most common being the production of siderophores, small iron-chelating molecules secreted into the bacterial environment. The opportunist pathogen Pseudomonas aeruginosa produces two siderophores, pyoverdine and pyochelin, and is also able to use a wide panoply of xenosiderophores, siderophores produced by other microorganisms. Here, we demonstrate that catecholamine neurotransmitters (dopamine, l-DOPA, epinephrine and norepinephrine) are able to chelate iron and efficiently bring iron into P. aeruginosa cells via TonB-dependent transporters (TBDTs). Bacterial growth assays under strong iron-restricted conditions and with numerous mutants showed that the TBDTs involved are PiuA and PirA. PiuA exhibited more pronounced specificity for dopamine uptake than for norepinephrine, epinephrine and l-DOPA, whereas PirA specificity appeared to be higher for l-DOPA and norepinephrine. Proteomic and qRT-PCR approaches showed pirA transcription and expression to be induced in the presence of all four catecholamines. Finally, the oxidative properties of catecholamines enable them to reduce iron, and we observed ferrous iron uptake via the FeoABC system in the presence of l-DOPA.

Lyte M.

More than 70 years ago, a new age in endocrinology was just beginning with the first purification of a hormone, adrenaline. As early as 1930, almost immediately following its first use, cases of adrenaline-associated sepsis were reported. From this time, there have been reports associating the elaboration of neuroendocrine hormones, such as adrenaline, with infectious disease. The most widely accepted theory to explain the ability of hormones to influence the course of infection involves the suppression of the immune system. The theory that the infectious microorganism itself might be equally responsive to the host's neuroendocrine environment has not been considered. It is the intent of this article to introduce a new perspective to the current understanding of the factors that mediate the ability of bacteria to cause disease, and to demonstrate that neuroendocrinology and microbiology intersect to form the interdisciplinary field of microbial endocrinology.

Park C.H., Valore E.V., Waring A.J., Ganz T.

Cysteine-rich antimicrobial peptides are abundant in animal and plant tissues involved in host defense. In insects, most are synthesized in the fat body, an organ analogous to the liver of vertebrates. From human urine, we characterized a cysteine-rich peptide with three forms differing by amino-terminal truncation, and we named it hepcidin (Hepc) because of its origin in the liver and its antimicrobial properties. Two predominant forms, Hepc20 and Hepc25, contained 20 and 25 amino acid residues with all 8 cysteines connected by intramolecular disulfide bonds. Reverse translation and search of the data bases found homologous liver cDNAs in species from fish to human and a corresponding human genomic sequence on human chromosome 19. The full cDNA by 5′ rapid amplification of cDNA ends was 0.4 kilobase pair, in agreement with hepcidin mRNA size on Northern blots. The liver was the predominant site of mRNA expression. The encoded prepropeptide contains 84 amino acids, but only the 20–25-amino acid processed forms were found in urine. Hepcidins exhibited antifungal activity against Candida albicans,Aspergillus fumigatus, and Aspergillus nigerand antibacterial activity against Escherichia coli,Staphylococcus aureus, Staphylococcus epidermidis, and group B Streptococcus. Hepcidin may be a vertebrate counterpart of cysteine-rich antimicrobial peptides produced in the fat body of insects.

Woods D.E., Jones A.L., Hill P.J.

Pseudomonas pseudomallei is the causative agent of melioidosis, a disease being increasingly recognized as an important cause of morbidity and mortality in many regions of the world. An intriguing observation regarding melioidosis is that a significant percentage of patients who develop the disease have preexisting diabetes mellitus. In this regard, we have tested the hypothesis that insulin may modulate the growth of P. pseudomallei. We have demonstrated that insulin markedly inhibits the growth of P. pseudomallei in vitro and in vivo. The growth rate of P. pseudomallei in minimal medium containing human recombinant insulin was significantly lower than that of control cultures containing no insulin. P. pseudomallei grew at an increased rate in serum samples obtained from diabetic rats compared with that in serum samples obtained from control animals. When the insulin level was restored by the addition of human recombinant insulin, the growth rate was reduced to a level similar to that seen in control serum. P. pseudomallei also grew significantly better in insulin-depleted human serum than control human serum. 125I-insulin binding studies demonstrated that P. pseudomallei possesses a specific, high-affinity binding site for human insulin. In in vivo studies, rats made diabetic by streptozotocin injection (80 mg/kg of body weight, intraperitoneally) were significantly more susceptible to P. pseudomallei septicemia than control rats. Thus, it appears that serum insulin levels may play a significant role in modulating the pathogenesis of P. pseudomallei septicemic infections.