1428 Trim29: tripartite motif-containing 29 Atf3: activating transcription factor 3 Mmp12: matrix metallopeptidase 12 Ahnak2: AHNAK nucleoprotein 2 Dnahc2: dynein, axonemal, heavy chain 2 Cdkn1c: cyclin-dependent kinase inhibitor 1C Mm.138637.1 Ms4a7: membrane-spanning 4-domains, subfamily A, member 7 Fabp5: fatty acid binding protein 5 9430019H13Rik: RIKEN cDNA 9430019H13 gene Msr1: macrophage scavenger receptor 1 Anpep: alanyl aminopeptidase Elane: elastase, neutrophil expressed F10: coagulation factor X Ms4a3: membrane-spanning 4-domains, subfamily A, member 3 Genes marked in bold are related to innate immunity and the genes marked in bold and also underlined are innate immunity genes catalogued by InnateDB. doi:10.1371/journal.pone.0048273.t003 order of their elevation in the brain, these are Lyz1, Lyz2, C1qb, Lgals3, C1qa, Grn, Ctss, Ctsd, Timp2, Man2b1, Hexb and Ctsb. Remarkably, other than Man2b1 and Hexb, the remaining ten are innate immune genes of which eight are lysosomal. All 12 may be putative, plasma predictors of the transition to cerebral disease. Elevation of LY-2835219 Lysozyme in BALB/c Npc1nmf164 Mice and its Reduction in Response to Treatment with Cyclodextrin, an Emerging Therapeutic Elevated Lysozyme Activity in the Plasma of Npc12/2 Mice Elevation of Innate Immunity in NPC Disease With the emergence of new therapeutics for NPC, there is urgent need for correlates whose levels mirror improvement of disease course as a consequence of treatment. Cyclodextrin has emerged as the most effective compound at retarding NPC disease in mice. Previous studies suggest that weekly injections of HPbCD to Npc1nih ameliorates the disease and extend the survival. Similarly, weekly injections of HPbCD to Npc1pf/pf mice also show improvement in disease status. We therefore treated Npc12/2 mice with HPbCD or vehicle control with once a week drug injections starting at age 2127 days. At 5055 days, untreated Npc12/2 mice had,1.41.8 fold higher plasma lysozyme activity compared to Npc1+/+ or Npc1+/2. The plasma lysozyme activity of the vehicle treated Npc12/2 mice remained elevated. However, it was significantly reduced in Npc12/2 mice treated with HPbCD. Thus, lysozyme may be an early disease correlate that measures responsiveness to a drug during the asymptomatic stage. Functional Validation of Elevated Innate Immunity Genes in Liver and Spleen of Npc12/2 Mice Elevation of Innate Immunity in NPC Disease Genes Entrez Gene 
IDLyz1: lysozyme 1 Lyz2: lysozyme 2 C1qb: complement component 1q, beta polypeptide Lgals3: Lectin, galactose binding, soluble3 C1qa: complement component 1q, alpha polypeptide Grn: granulin Ctss: cathepsin S Ctsd: cathepsin D Timp2: tissue inhibitor of metalloproteinase 2 Man2b1: mannosidase 2, alpha B1 Hexb: hexosaminidase B Ctsb: cathepsin B Genes marked in bold code for secretory lysosomal proteins. doi:10.1371/journal.pone.0048273.t004 bacterial loads in Npc1+/+ and Npc1+/2 mice. However, there was,810 fold reduction in bacterial load in the organs of Npc12/2 mice. Since the spleen is readily amenable to comprehensive cellular analysis of innate immunity, we examined the numbers of CD335+ natural killer cell, CD11c+ dendritic cells, CD11b+F4/ 80+ monocytes and macrophages, and CD11b+Gr-1hi neutrophils in splenic single cell suspensions of Npc12/2 and Npc1+/2 animals. Again, we selected mice of age at 68 weeks, because the reason described above. Flow cytometric 15647369 target=_blank”>9874164 analysis showed no effect on counts of NK cell or dendritic cel
