In the original article, there was a mistake in Figure 8A, as published. We showed that we did not get expression of the FLaB1 subunit in Manilae L495 strain. In fact, the forward primer (designed according to the Fiocruz sequence) used to amplify the FlaB1 subunit has two mismatches within the Manilae sequence. We did the RT-PCR with the good primer and found an amplification, showing an enhanced expression at the stationary phase compared to the exponential phase, likewise the other FlaB subunits. The corrected Figure 8 appears below.
Figure 8.
(A) In vitro FlaBs mRNA expression in L. interrogans Copenhageni Fiocruz L1-130, Icterohaemorrhagiae Verdun and Manilae L495 at the exponential (E) and stationary (S) phase. Data of RT-qPCR are expressed as the relative mRNA quantities normalized to the expression of the lipl41 mRNA. Technical replicates are represented as dots and lines correspond to mean (+/- SD) of replicates (3 < n < 9). Statistically significant differences (Student t-test) are indicated. (B) In vivo FlaAs and (C) FlaBs mRNA expression in blood of infected mice (n=5, light blue) and hamsters (n-5, dark blue), 24 h post intraperitoneal infection with 2x108 virulent L. interrogans Icterohaemorrhagiae strain Verdun, compared with mRNA expression in culture in EMJH at 30° C. Data of RT-gPCR are expressed as the ratio of mRNA quantities relatives to the EMJH control. Individual animals are represented as dots and lines correspond to mean (+/- SD) of all animals. Statistically significant differences (Student t-test) are indicated with corresponding p values: * for p < 0.05; ** for p < 0.01 and *** for p < 0.001.
A correction has been made to Results, “FlaB mRNA Are Upregulated in Stationary Phase”.
“Of note, and different from other strains, the Manilae L495 flaB1 mRNA was undetectable” and a paragraph in the Discussion, “Furthermore, in Manilae L495…” are not accurate and should be both omitted.
In the original article, there was also mistake in Figure 6, Supplementary Figure 3, Supplementary Figure 4 and Supplementary Figure 6 and their legends as published. We realized a “slipping” of the names of FlaB subunits only in the figures of alignments and comparison of sequences (Figure 6, Sup Figure 3, Sup Figure 4 and Sup Figure 6). What we called FlaB1 in figures and respective legends is in fact FlaB4, FlaB2 is FlaB1, FlaB3 is FlaB2, and FlaB4 is FlaB3 (see corrected annotation in Additional table provided).
Figure 6.
(A) Amino acid sequence homology average percentage between Salmonella typhimurium FliC (P06179) and Leptospira interrogans strain Fiocruz FlaB (LIC11890, LIC11889, LIC 11532 and LIC11531) and FlaAs (LIC10788 and LIC10787) and primary structures of the flagellin proteins with TLR5 binding consensus. (B) In silico (Phyre2 and Chimera softwares) prediction of Salmonella typhimurium FliC (P06179) structure with the four described domains and with positions of the TLR5 binding consensus: 1 (red), 2 (yellow) and 3 (light blue) and stabilization region (light green) highlighted. (C) In silico (Phyre2 and Chimera softwares) prediction of Leptospira interrogans strain Fiocruz FlaB4 (LICI1531) with the positions of the TLRS binding consensus and stabilization region highlighted, FlaA1 (LIC10788), FlaA2 (LICI0787). (D) Clustal (MEGA software) alignment of the amino acid sequences for the TLR5 binding consensus regions of: Salmonella enterica FliC (GeneBank QDQ31983.1), L. bifleva (strain Patoc) FlaB1 (LEPBla2133), FlaB2 (LEPBIa2132), FlaB3 (LEPBla1872) and FlaB4 (LEPBla1589), L. interrogans (strain Fiocruz L1-130) FlaB1 (LIC18890), FlaB2 (LICIT889), FlaB3 (LICI1532) and FlaB4 (LIC11531), L. interrogans (strain L495) FlaB1 (LMANv2 260016), FlaB2 (LMANv2 260015). FlaB3 (LMANv2 590024) and FlaB4 (LMANv2 590023), and L. interrogans (strain Verdun) FlaB1 (AKWP_v1_110429), FlaB2 (AKWP_v1_110428) and FlaB3 (AKWP_v1_110068) and FlaB4 (AKWP_v1_110067).
New annotations consistent with studies from M Picardeau.
| 10788 | Patoc* | Fiocruz L1-130* | Manilae* | Verdun* |
|---|---|---|---|---|
| flaA1 | LEPBIa2335 | LIC10788 | LMANv2_260046 / LIMLP_13775 | AKWP_v1_210009 |
| flaA2 | LEPBIa2336 | LIC10787 | LMANv2_260045 / LIMLP_13780 |
AKWP_v1_210008 |
| flaB1 | LEPBIa2133 | LIC11890 | LMANv2_260016 / LIMLP_09410 |
AKWP_v1_110429 |
| flaB2 | LEPBIa2132 | LIC11889 | LMANv2_260015 /LIMLP_09405 |
AKWP_v1_110428 |
| flaB3 | LEPBIa1872 | LIC11532 | LMANv2_590024 / LIMLP_07480 |
AKWP_v1_110068 |
| flaB4 | LEPBIa1589 | LIC11531 | LMANv2_590023 / LIMLP_07475 |
AKWP_v1_110067 |
*Annotation of L. biflexa serovar Patoc strain Patoc 1 (Patoc), L. interrogans serovar Copenhageni strain Fiocruz L1-130 (Fiocruz L1-130), L. interrogans serovar Manilae strain L495 (Manilae), L. interrogans serovar Manilae UP-MMC-NIID LP (Manilae) (ref Satou et al. 2015 ; PMID: 26272567).
L. interrogans serovar Icterohaemorrhagiae Verdun LP (Verdun) in https://mage.genoscope.cns.fr/microscope/home/index.php.
Additional Table Old annotations (published Frontiers Immunol 2020).
| 10788 | Patoc | Fiocruz L1-130 | Manilae | Verdun |
|---|---|---|---|---|
| flaA1 | LEPBIa2335 | LIC10788 | LMANv2_260046 | AKWP_v1_210009 |
| flaA2 | LEPBIa2336 | LIC10787 | LMANv2_260045 | AKWP_v1_210008 |
| flaB1 | LEPBIa1589 | LIC11531 | LMANv2_590023 | AKWP_v1_110067 |
| flaB2 | LEPBIa2133 | LIC11890 | LMANv2_260016 | AKWP_v1_110429 |
| flaB3 | LEPBIa2132 | LIC11889 | LMANv2_260015 | AKWP_v1_110428 |
| flaB4 | LEPBIa1872 | LIC11532 | LMANv2_590024 | AKWP_v1_110068 |
However, the annotations used in Figures 7, 8 were correct and all the mutants used are correct. The corrected Figure 6 and its corrected legend appears below. The corrected Supplementary Figures 3, 4, and 6 can be accessed from the original article.
The authors apologize for these errors and state that they do not change the scientific conclusions of the article in any way. The original article has been updated.
Publisher’s note
All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.