Biomarkers showed different responses to standard immunosuppressant therapy.

<p>Biomarkers showed different responses to standard immunosuppressant therapy.</p

Monocyte Chemotactic Protein-1, Fractalkine, and Receptor for Advanced Glycation End Products in Different Pathological Types of Lupus Nephritis and Their Value in Different Treatment Prognoses

<div><p>Background</p><p>Early diagnosis is important for the outcome of lupus nephritis (LN). However, the pathological type of lupus nephritis closely related to the clinical manifestations; therefore, the treatment of lupus nephritis depends on the different pathological types.</p><p>Objective</p><p>To assess the level of monocyte chemotactic protein (MCP-1), fractalkine (Fkn), and receptor for advanced glycation end product (RAGE) in different pathological types of lupus nephritis and to explore the value of these biomarkers for predicting the prognosis of lupus nephritis.</p><p>Methods</p><p>Patients included in this study were assessed using renal biopsy. Class III and class IV were defined as the proliferative group, class V as non-proliferative group, and class V+III and class V+IV as the mixed group. During the follow-up, 40 of 178 enrolled patients had a poor response to the standard immunosuppressant therapy. The level of markers in the different response groups was tested.</p><p>Results</p><p>The levels of urine and serum MCP-1, urine and serum fractalkine, and serum RAGE were higher in the proliferative group, and lower in the non-proliferative group, and this difference was significant. The levels of urine and serum MCP-1 and serum RAGE were lower in the poor response group, and these differences were also significant. The relationship between urine MCP-1 and urine and serum fractalkine with the systemic lupus erythematosus disease activity index was evaluated.</p><p>Conclusion</p><p>The concentration of cytokines MCP-1, fractalkine, and RAGE may be correlated with different pathology type of lupus nephtitis. Urine and serum MCP-1 and serum RAGE may help in predicting the prognosis prior to standard immunosuppressant therapy.</p></div

Baseline status of proliferative, non-proliferative, and mixed groups.

<p>Baseline status of proliferative, non-proliferative, and mixed groups.</p

MCP-1, fractalkine and RAGE had different levels for the different pathological types of lupus nephritis.

<p>MCP-1, fractalkine and RAGE had different levels for the different pathological types of lupus nephritis.</p

Level of biomarkers in the good response to immunosuppressant therapy group and the poor response to immunosuppressant therapy group.

<p>Level of biomarkers in the good response to immunosuppressant therapy group and the poor response to immunosuppressant therapy group.</p

The GLS is required for <i>gurken</i> RNA localization and gene function.

<p>(A–B) Wild-type expression patterns of endogenous <i>gurken</i> RNA (A) and protein (B) as revealed by whole mount <i>in situ</i> hybridization and immunofluorescence, respectively. Anterodorsal localization of transcripts and protein is only apparent in the rightmost egg chambers, which are stage 8 and 9, respectively. (C–E) The <i>gurken</i> RNA and protein distribution patterns of <i>gurken</i> null mutants (<i>grk^ΔFRT</i>) carrying the wild-type <i>gurken</i> transgene, <i>grk^wt</i> (C–D) or no transgene (E). (F–H) <i>grk^ΔFRT</i> eggs and egg chambers (from <i>gurken</i> null mothers) carrying the <i>grkGLS^mut</i> transgene. (F) Left panel: representative <i>grk^ΔFRT</i>; <i>grkGLS^mut</i> egg exhibiting a completely ventralized phenotype, i.e., complete loss of dorsal appendage material. Right panel; anterior end of a <i>grk^ΔFRT</i>; <i>grkGLS^mut</i> egg exhibiting a strong, but not complete, ventralized phenotype. Note, for example the short, fused dorsal appendage. (G) <i>grk^ΔFRT</i>; <i>grkGLS^mut</i> ovariole following <i>in situ</i> hybridization with <i>gurken</i> probe. Transcripts are dispersed throughout the germ-line cysts with only slight enrichment in the oocyte and no subcellular localization. (H) <i>grk^ΔFRT</i>; <i>grkGLS^mut</i> ovariole following immunofluorescence using an anti-Grk antibody. The protein is generally dispersed throughout the germ-line cysts, although slight enrichment around the oocyte nucleus is seen in rare stage 10 and 11 egg chambers.</p

Conservation and predicted secondary structure of the GLS.

<p>(A) Sequence alignment of the <i>gurken</i> transcription unit displayed using the Vista Browser at <a href="http://pipeline.lbl.gov/cgi-bin/gateway2" target="_blank">http://pipeline.lbl.gov/cgi-bin/gateway2</a><a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0015448#pone.0015448-Nielsen1" target="_blank">[49]</a>. The estimated years in millions (MYA) of evolution between <i>D</i>. <i>melanogaster</i> and each of the other five species is from Heger and Ponting <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0015448#pone.0015448-Heger1" target="_blank">[24]</a>. The most highly conserved region is circled and includes the first 39 nt of the GLS. The last 25 nt of the GLS map to the 3′ side of the abutting intron. The arrow indicates the direction of transcription. The red shaded region corresponds to a putative transposable element. The numbers at the bottom of the graph indicate nucleotide position along the chromosome. (B) The 5′ end of the <i>gurken</i> mRNA, where the green dot denotes the translation start site, the red arrows the boundaries of the GLS, and the asterisk the position of the intron. The nucleotides beneath the aligned sequence blocks highlight differences between the <i>D. Willistoni</i> and <i>D. melanogaster</i> sequences. (C) Predicted secondary structure of the GLS, with non-conserved residues shown in red.</p

The baseline status of different response to the immune therapy.

<p>The baseline status of different response to the immune therapy.</p

Structure of GLS variants.

<p>The wild-type GLS is shown at the left for comparison. The GLS mutant (referred to as <i>grkGLS^mut</i> in Text) contains 12 point mutations (shown in red), which are predicted to disrupt the predicted base pairing pattern of the GLS at five sites (circled). None of the 12 mutations affect the protein coding sequence as shown at the bottom portion of the figure.</p

Rab11 is required for the survival of follicle epithelial cells.

<p>(A–E) Confocal images of immunostained germaria and/or egg chambers 10–12 days ACI, with <i>rab11-null</i> cells marked by the absence of nGFP. All egg chambers are derived from mosaic germaria that contained one <i>rab11-</i>null FSC and one wildtype FSC as evidenced by direct examination of the adjacent germarium and/or the presence of <i>rab11-null</i> cells in the egg chambers themselves. The stages of the compound and fused egg chambers are estimates based on the size of the nurse cell nuclei, the distance of the fused/compound egg chamber from the germarium, and/or its position relative to other more easily staged egg chambers within the same ovariole. (A) An s5 egg chamber fused to a compound egg chamber containing two ∼s6/7 germline cysts encased in a single continuous epithelium and immunostained for nGFP (green) and E-cad (red). <i>rab11-null</i> cells, which are marked with asterisks, are more abundant in the s5 egg chamber than in the adjacent older ones. The positions of the <i>rab11-null</i> cells in the compound egg chamber are consistent with a stalk or stalk-like identity (discussed more fully in Text and see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0020180#pone-0020180-g003" target="_blank">Fig. 3</a>). (B) An s5 egg chamber fused to an s8 or s9 egg chamber immunostained for nGFP (Green) and Fas3 (red). The arrowhead points to a cluster of 2–3 <i>rab11-null</i> cells that over express Fas-3. While such expression is indicative of a polar cell fate, we cannot rule out a stalk cell fate for these cells as stalk cells that fail to incorporate themselves into a functional stalk are also known to over-express Fas3 <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0020180#pone.0020180-AssaKunik1" target="_blank">[43]</a>. No <i>rab11-null</i> epithelial cells are seen in this plane of focus. Other focal planes (not shown) contained no or only a few <i>rab11-null</i> epithelial cells (not shown). The two strongly expressing Fas3 cells at the posterior end of the s8/9 egg chamber are wildtype polar cells, although the GFP signal is weak at the focal plane shown. (C) Germarium and s3 egg chamber immunostained for nGFP (Green) and E-cad (red). The arrow points to a putative wildtype FSC. A large <i>rab11-null</i> clone in the germarium is outlined in white. The dashed yellow line in the adjacent s3 egg chamber highlights two large gaps in the epithelium as evident by the absence of E-cad expression and also Nomarski imaging (not shown). (D) Germarium and s2 and s3 egg chambers immunostained for nGFP (green) and counterstained for activated caspase 3, a marker for PCD. Approximately half of the <i>rab11-null</i> cells (outlined with the dashed line) in the s3 egg chamber stain positively for activated caspase 3. The apparent weaker staining of some of the <i>rab11-null</i> cells is not evident at other focal planes (not shown). The activated caspase 3 staining activity at the junction between germarial regions 1 and 2a (arrowhead) is likely to correspond to escort cells, which are known to be targeted for PCD at this stage <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0020180#pone.0020180-Decotto1" target="_blank">[44]</a>. (E) Control <i>rab11-null</i> ; P[<i>rab11+</i>] clones in s4 and s7 egg chambers immunostained for nGFP (green), E-cad (red).</p