"<i>Candidatus</i> Borrelia kalaharica" Detected from a Febrile Traveller Returning to Germany from Vacation in Southern Africa - Fig 2

<p><b>Molecular phylogenetic analysis using the maximum likelihood method based on partial sequences of (A) 16S rRNA (473 bp), (B) <i>flaB</i> (694 bp) and (C) <i>uvrA</i> (900 bp)</b>. The taxa labels show the NCBI accession number, <i>Borrelia</i> species, and strain information (if available). The strain investigated in this study (indicated by a black dot)–although acquired in Africa–clusters more closely to New World RF species in <i>flaB</i> (B) and <i>uvrA</i> (C) phylogenies. The trees with the highest log likelihood are shown. The percentage of trees in which the associated taxa clustered together is shown next to the branches. Scale bar: substitutions per site. RF = relapsing fever; LB = Lyme borreliosis</p

Microphotograph of a thick blood smear of the patient at the time of admission.

<p>The image shows spirochetes together with leucocytes. Striking is the relative short length of the bacterial cells (10μm) and the relative small number of turns. This morphology was observed in all intact bacteria in the slide. Pictures were taken with a 100x oil immersion objective and a T2 DSLR photo adapter.</p

Estimates of evolutionary divergence between 16S rRNA sequences.

<p>The number of base substitutions per site from between sequences are shown. Analyses were conducted using the Kimura 2-parameter model. There were a total of 473 positions in the final dataset.</p

Estimates of evolutionary divergence between <i>flaB</i> sequences.

<p>The number of base substitutions per site from between sequences are shown. Analyses were conducted using the Kimura 2-parameter model. The analysis involved 18 nucleotide sequences. There were a total of 640 positions in the final dataset.</p

Estimates of evolutionary divergence between <i>uvrA</i> sequences.

<p>The number of base substitutions per site from between sequences are shown. There were a total of 900 positions in the final dataset.</p

Baseline Characteristics^*.

<p>*presented as median values.</p><p>**nucleosid/nucleotide analogs.</p><p>***not determined.</p

Effect of PD-L1/2 blockade on CD4⁺ T-cell expansion.

<p>Induction of CD4⁺ T-cell proliferation in chronic HBV (n = 23) from h0 (<i>left</i>) to antigenic stimulation (<i>middle</i>) and PD-L1/2 blockade (<i>right</i>) illustrated as point to point graphs from (<b>A</b>) PD-L1/2 responders (n = 9) and (<b>B</b>) Non-responders (n = 14). Contour plots are shown for each group. (<b>C</b>) Patients treated with nucleosid/nucleotid analogs (<i>black bars</i>) responded to PD-1 blockade using Chi²-test, indicating that viral control let enhance patients susceptibility as (<b>D</b>) treated patients are characterized by low viremia. (<b>E</b>) Decreased PD-1 expression in PD-L1/2 responders (R) (n = 6) versus non-responders (NR) (n = 6). (<b>F</b>) CD4⁺ T-cell frequencies upon antigenic stimulation (<i>black bars</i>) and PD-L1/2 inhibition (<i>grey bars</i>) in relation to PD-1 expression.</p

Inhibitory phenotype of HBV-specific CD4⁺ T-cells.

<p>(<b>A</b>) PD-1 (n = 29), CTLA-4 (n = 8), TIM-3 (n = 14), KLRG1 (n = 12) and CD244 (n = 8) expression is displayed on Tetramer⁺CD4⁺ T-cells during chronic HBV. (<b>B</b>) PD-1 expression in chronic (<i>black points</i>) (n = 29) and acute HBV (<i>black squares</i>) (n = 7), resolved HBV (<i>white squares</i>) (n = 5), EBV (<i>grey points</i>) (n = 15) and Flu (<i>white points</i>) (n = 21) infection. (<b>C</b>) Contour plots illustrate patterns of PD-1 expression during HBV, EBV and Flu infection. (<b>D</b>) CD127 expression on HBV-specific (<i>black points</i>) (n = 11) and Flu-specific CD4⁺ T-cells (<i>white points</i>) (n = 9). (<b>E</b>) Contour plots display CD127 expression on CD14⁻, CD19⁻, Via Probe⁻, Tetramer⁺ T-cells. (<b>F</b>) CD127 defines PD-1 expression as examined in HBV (<i>black bars</i>) and Flu infection (<i>white bar</i>). (<b>G</b>) Influence of viral load on CD127 expression.</p

Effect of PD-L1/2 blockade on cytokine release.

<p>Analysis of cytokine⁺CD4⁺ T-cells allows to differ (<b>A</b>) PD-L1/2 responders (n = 4) from (<b>B</b>) Non-responders (n = 9). Floating bars illustrate mean values ranging from minimun to maximum production of IFN-γ, IL-2 and TNF-α seperatly following antigen stimulation (<i>white bars</i>) and PD-1 blockade (<i>grey bars</i>). Contour plots are shown for IFN-γ (<i>top</i>), IL-2 (<i>middl</i>e) and TNF-α (<i>bottom</i>) secretion upon antigen stimulation (<i>left</i>) and PD-1 pathway neutralization (<i>right</i>) after gating on CD3⁺CD4⁺ T-cells.</p