Background
Methods
Descriptions of mice and T. gondii infection of mice for behavioral and neurologic assessments, MRIs, microarrays, and histopathology
Swiss Webster mice
Viral pathogens
Bacterial pathogens
Parasitic pathogens
Infection with T. gondii
Experimental design/cohorts studied
Descriptions of mice for histopathologic studies
Behavioral assessments
Physical appearance and autonomic characteristics
Illness behavior
Hunching
Piloerection
Grooming/Fur
Palpebral closure
Salivation
Urination/Defecation
Tail wounds
Body position/posture
Body weight
Tremor
Activity
Gait
Transfer arousal
Spatial locomotion
Stereotyped behavior
Exploratory behavior
Open-field testing
Neuromuscular function
Grip strength
Vertical pole test
Sensorimotor function
Pain insensitivity
MRI
Gene expression and statistical analysis of microarrays
Histopathology
Immunohistochemistry and electron microscopy
Immunostaining for T cells and activated microglia
Immunostaining for tachyzoites and bradyzoites and identification of host plasma cells
Electron microscopy
Myelin and other special neuronal staining and histopathology
Lipoprotein and SAA analysis
Knockout mice
Chronically infected and uninfected control Balb/c mice
Treatment of mice with sulfadiazine
Treatment of mice with hamster polyclonal antibody to PD-L1 or hamster isotype-control
Statistical analysis of behavioral and neurologic findings
Statistical analysis of histopathologic findings
Results
Appearance, behavior, and neurological findings in the initial cohort
APPEARANCE | LOCOMOTION | ANS | OPEN FIELD ACTIVITY | |||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Condition | Grooming | Body Position | Piloerection | Tail Damage | Tremor | Palpebral Closure | Lacrimation | Weight (gm) | Spatial Locomotion | Gait | Urination/Defecation | Sniffs | Rears | Center | Freezing | Latency to move (sec) |
Control | Good | - | - | - | - | - | - | 39.4 | Active | - | - | 2 | 20 | 4 | 0 | 4 |
Control | Good | - | - | - | - | - | - | 52.3 | Active | - | - | 4 | 4 | 2 | 0 | 22 |
Control | Good | - | - | - | - | - | - | 45.0 | Active | - | - | 6 | 13 | 3 | 0 | 41 |
Control | Good | - | - | - | - | - | - | 50.5 | Active | - | - | 1 | 0 | 7 | 0 | 120 |
Control | Good | - | - | - | - | - | - | 50.6 | Active | - | - | 4 | 9 | 7 | 0 | 10 |
Control | Good | - | - | - | - | - | - | 58.5 | Active | - | - | 1 | 2 | 9 | 0 | 1 |
Infected | Poor | + | + | + | + | - | - | 43.8 | Slow | + | + | nd | nd | nd | nd | nd |
Infected | Good | + | + | + | - | - | - | 41.2 | Active | + | + | 0 | 0 | 0 | 0 | 180 |
Infected | Very Poor | + | - | + | + | - | - | 42.0 | None | + | + | 0 | 0 | 0 | 0 | 180 |
Infected | Very Poor | + | + | - | + | + | + | 35.3 | None | + | + | nd | nd | nd | nd | nd |
Infected | Good | - | + | + | - | - | - | 34.9 | Active | - | + | 0 | 0 | 0 | 0 | 180 |
Infected | Very Poor | + | + | - | + | + | + | 34.8 | Slow | + | + | 0 | 0 | 0 | 0 | 180 |
Infected | Poor | + | + | + | + | + | + | 35.6 | Slow | + | + | 1 | 3 | 1 | 1 | 24 |
Infected | Sub-normal | + | + | - | + | + | + | 36.2 | Active | - | + | 0 | 2 | 10 | 1 | 6 |
Infected | Very Poor | + | + | + | - | - | + | 58.0 | None | - | + | 0 | 0 | 0 | 0 | 180 |
Infected | Poor | + | + | + | + | - | - | 54.2 | Slow | - | + | 2 | 0 | 1 | 2 | 56 |
P-values | 0.018 | 0.001 | 0.001 | 0.011 | 0.011 | 0.23 | 0.093 | 0.059 | 0.035 | 0.034 | < 0.001 | 0.0055 | 0.016 | 0.017 | 0.11 | 0.021 |
Kinetics of development of behavioral and neurologic abnormalities in two additional studies (replicates of each other)
Brain MRIs of chronically infected mice have mild to moderate ventricular dilatation
Correlation of brain weight and neurologic findings
Microarrays reflect inflammation and increased expression of CD36 and PD1L, GFAP, ubiquitin ligase, and C1q
Function | Gene* [Number/ID]† |
---|---|
Antibody | Ig heavy chain V regions [116] |
Ig kappa V regions [119] | |
Ig lambda 1 germline V region (Igl-V1, 1810027O01Rik, IgL) [V00811] | |
Ig alpha chain (IGHV1S44) [M19402] | |
Rearranged IgA-chain gene, V region (IGHV1S46) [M20774] | |
IgG-1 gene, D-J-C region: 3' exon for secreted form (Igh-4, IgG1) [J00453] | |
Ig germline D-J-C region alpha gene and secreted tail (Igh-VJ558, AI893585, MGC118142, MGC6727, Igh-A (1g2), Vh186.2/Jh2) [J00475] | |
Ig gamma2a-b(c57bl/6 allele) c gene and secreted tail (IGHG2C) [J00479] | |
Immunoglobulin joining chain (Igj, 9530090F24Rik, AI323815, Jch) [NM_152839] | |
Antigen presentation | MHC-I Similar to H-2 MHC-I antigen, D-37 alpha chain precursor (C920025E04Rik) [AK083387] |
MHC-I Transporter 2, ATP-binding cassette, sub-family B (MDR/TAP) (Tap2, ABC18, AI462429, APT2, Abcb3, Ham-2, Ham2, MTP2, PSF2, RING11, Tap-2, Y1, jas) [NM_011530] | |
MHC-II [22] | |
MHC-III (D17H6S56E-3, C6orf27, G7c, NG37) [NM_138582] | |
Beta-2 microglobulin (B2m, Ly-m11, beta2-m) [NM_009735] | |
Complement | Complement component 4A/sex limited protein (C4a, Slp) GeneCards: Primarily expressed in liver and to a lesser extent in immune cells [NM_011413] |
Complement component 4B (C4b, C4, Ss) [NM_009780] | |
Complement component 1, r subcomponent (C1r, AI132558, C1rb) [NM_023143] | |
Complement component C1SB (C1sb) [NM_173864] | |
Complement component 1, q subcomponent, alpha polypeptide (C1qa, AI255395, C1q) [NM_007572] | |
Complement component 1, q subcomponent, C chain (C1qc, AI385742, C1qg, Ciqc) [NM_007574] | |
Major inhibitor of classical complement | Serine (or cysteine) peptidase inhibitor, clade G, member 1 (Serping1, C1INH, C1nh) [NM_009776] |
Serine (or cysteine) peptidase inhibitor, clade A, member 3N (Serpina3n, Spi2-2, Spi2.2, Spi2/eb.4) GeneCards: Although its physiological function is unclear, it can inhibit neutrophil cathepsin G and mast cell chymase. [NM_009252] | |
GTPase | Interferon inducible GTPase 1 (Iigp1, 2900074L10Rik, AI046432, AW111922, Iigp) [AK013785] |
Interferon inducible GTPase 1 (AW111922, Iigp1) [NM_021792] | |
Interferon inducible GTPase 2 (Iigp2, RP24-499A8.1, AI481100, GTPI, (GC102455) [NM_019440] | |
Interferon gamma inducible protein 47 (Ifi47, IRG-47, Iigp4, Iipg4, Olfr56) [NM_008330] | |
Interferon gamma induced GTPase (Igtp, RP24-499A8.4, AW558444) [NM_018738] | |
GTPase, IMAP family member 4 (Ian1, Gimap4, AU019574, E430007K16Rik, IMAP4, MGC11734, mIAN1), transcript variant 1 GeneCards: Exhibits intrisinic GTPase activity. [NM_174990] | |
Transcription | Signal transducer and activator of transcription (Stat1, 2010005J02Rik, AA408197) GeneCards: Mediates signaling by interferons. Results in induction of a cellular antiviral state. [NM_009283 BC057690] |
Transcription factor MafB (v-maf musculoaponeurotic fibrosarcoma oncogene homolog B).(Mafb, Kreisler, Krml, kr) GeneCards: Plays a pivotal role in regulating lineage-specific hematopoiesis by repressing ETS1-mediated transcription of erythroid-specific genes in myeloid cells (By similarity). [NM_010658] | |
Down regulator of transcription 1/Differentially regulated in lymphoid organs and differentiation (Dr1d) [AF043513] | |
Suppression of cytokine signaling | Suppressor of cytokine signaling 1 (Socs1, Cish1, Cish7, JAB, SOCS-1, SSI-1) GeneCards: SOCS1 is involved in negative regulation of cytokines that signal through the JAK/STAT3 pathway. Appears to be a major regulator of signaling by interleukin 6 (IL6) and leukemia inhibitory factor (LIF). Regulates interferon-gamma mediated sensory neuron survival (By similarity). [NM_009896] |
Proteosome/antigen processing | Proteosome (prosome, macropain) subunit, beta type 8 (large multifunctional peptidase 7) (Psmb8, Lmp-7, Lmp7) GeneCards: This subunit is involved in antigen processing to generate class I binding peptides. Stimulated by interferon gamma, involved in the degradation of cytoplasmic antigens for MHC class I antigen presentation pathways. [NM_010724] |
Proteosome (prosome, macropain) subunit, beta type 9 (large multifunctional peptidase 2) (Psmb9, Lmp-2, Lmp2) GeneCards: stimulated by interferon gamma, involved in the degradation of cytoplasmic antigens for MHC class I antigen presentation pathways. [NM_013585] | |
Proteasome (prosome, macropain) 28 subunit, alpha (Psme1, AW413925, MGC113815, PA28a) GeneCards: Implicated in immunoproteasome assembly and required for efficient antigen processing. The PA28 activator complex enhances the generation of class I binding peptides by altering the cleavage pattern of the proteasome. Induction by interferon gamma. [NM_011189] | |
Antimicrobial | Secretory leukocyte peptidase inhibitor (Slpi) Acid-stable proteinase inhibitor with strong affinities for trypsin, chymotrypsin, elastase, and cathepsin G. Secretory leukoprotease inhibitor, involved in antineutrophil elastase protection at inflammatory sites. [NM_011414] |
Defensin related cryptdin 17 (Defcr17, AU014719, Cryp17) GeneCards: Acid-stable proteinase inhibitor with strong affinities for trypsin, chymotrypsin, elastase, and cathepsin G. Secretory leukoprotease inhibitor, involved in antineutrophil elastase protection at inflammatory sites [S73391] | |
Pre-B-cell growth | Bone marrow stromal cell antigen 2 (Bst2, 2310015I10Rik, C87040, DAMP-1) GeneCards: May be involved in pre-B-cell growth. [NM_198095] |
T lymphocyte proliferation | CD274 antigen/Programmed cell death 1 ligand 1 precursor (Cd274, B7-H1, PD-L1, Pdcd1l1, Pdcd1lg1) Genecards: Involved in costimulatory signal, essential for T lymphocyte proliferation and production of IL10 and IFNG, in an IL2-dependent and a PDCD1-independent manner. Interaction with PDCD1 inhibits T-cell proliferation and cytokine production. Up-regulated on T and B cells, dendritic cells, keratinocytes and monocytes after LPS and IFNG activation. Up-regulated in B cells activated by surface Ig cross-linking. [NM_021893] |
Lymphocyte activation | Lymphocyte-activation gene 3 (Lag3, CD223, LAG-3, Ly66) GeneCards: Involved in lymphocyte activation. Binds to HLA class-II antigens. Highly homologous to CD4, expressed exclusively in activated T and NK lymphocytes, major MHC class 2 ligand potentially involved in the regulation of immune response. [NM_008479] |
Macrophage growth | Guanylate nucleotide binding protein 2 (Gbp2) Uniprot: Interferon-induced Gbp 2 [NM_010260] |
Guanylate nucleotide binding protein 4 (Gbp4, AW228655, Gbp3) [NM_018734] | |
Macrophage activation 2/guanylate-binding protein 4 homolog (Mpa2, AW228052, Gbp4, KIAA4245, Mag-2, Mpa-2, mKIAA4245) [NM_008620] | |
Macrophage activation 2 like (MPA2l, AI595338) [NM_194336] | |
Guanylate nucleotide binding protein 5 (Gbp5, 5330409J06Rik) Uniprot: Interferon-induced Gbp5 [NM_153564] | |
RIKEN cDNA 5830443L24 gene/guanylate binding protein 8( 5830443L24Rik, mGBP8) [NM_029509] | |
cDNA sequence BC057170/guanylate binding protein like (BC057170, E430029F06) [NM_172777] | |
Immune response/binding | Interferon-induced protein with tetratricopeptide repeats 3 (Ifit3, Ifi49, MGC107331) [NM_010501] GO:0000004 biological process unknown GO:0005488 binding GO:0005554 molecular function unknown GO:0006955 immune response GO:0008372 cellular component unknown |
Astrocyte protein after trauma | Glial fibrillary acidic protein (Gfap, AI836096) GeneCards: Class-III intermediate filament, cell-specific marker; during development of CNS distinguishes astrocytes from other glial cells. Almost exclusively expressed in astrocytes; interacts with S100A1. S100A1 home page: S100A1 and S100B can be isolated as a complex from bovine brain; mixture of S100A1 and S100B inhibited the assembly of tubulin into microtubules [NM_010277] |
Chemo-attractants | Chemokine (C-C motif) ligand 5 (Ccl5, MuRantes, RANTES, SISd, Scya5, TCP228) GeneCards: Chemoattractant for blood monocytes, memory T helper cells and eosinophils. [NM_013653] |
Chemokine (C-C motif) ligand 8/monocyte chemoattractant protein-2 precursor (Ccl8, RP23-446K18.1, 1810063B20Rik, AB023418, HC14, MCP-2, Mcp2, Scya8) GeneCards: Interferon gamma induced chemotaxis Attracts monocytes, lymphocytes, basophils, eosinophils [NM_021443] | |
Cell adhesion/interaction | CD36 antigen/fatty acid translocase (Cd36, FAT, GPIV, Scarb3) May function as a cell adhesion molecule. GeneCards: Directly mediates cytoadherence of Plasmodium falciparum parasitized erythrocytes. Mediates free radical production in cerebral ischemia. Publication (Khoury, J Exp Med 2003, 197(12) 1657–1666): CD36, a major pattern recognition receptor, mediates microglial and macrophage response to beta-amyloid, and imply that CD36 plays a key role in the proinflammatory events associated with Alzheimer's disease. [NM_007643] |
EGF-like module containing, mucin-like, hormone receptor-like sequence 1/cell surface glycoprotein F4/80/lymphocyte antigen 71 (Emr1, D7A5-7, EGF-TM7, F4/80, Gpf480, Ly71, TM7LN3) GeneCards: Could be involved in cell-cell interactions. [NM_010130] | |
Lectin, galactoside-binding, soluble, 3 binding protein/cyclophilin associated. protein (Lgals3bp, 90K, CyCAP, MAC-2BP, Ppicap) GeneCards: Promotes integrin-mediated cell adhesion. May stimulate host defense against viruses and tumor cells. [NM_011150] | |
Leukocyte integrin/actin signaling | FYN binding protein (Fyb, ADAP, B630013F22Rik) GeneCards: Acts as an adapter protein of the FYN and SH2-domain-containing leukocyte protein-76 (SLP76) signaling cascades in T cells. Modulates expression of interleukin-2 (IL-2). [NM_011815] |
Free radical production | Cytochrome b-245, beta polypeptide (Cybb, C88302, Cgd, Nox2, gp91<phox>, gp91phox) GeneCards: Critical component of the membrane-bound oxidase of phagocytes that generates superoxide. [NM_007807] |
Apoptosis retardation | B-cell leukemia/lymphoma 2 related protein A1c(Bcl2a1, A1-c c) GeneCards: Retards apoptosis induced by IL-3 deprivation. May function in response of hemopoietic cells to external signals and in maintaining endothelial survival during infection (By similarity). [NM_007535] |
ADP ribosylation | Poly (ADP-ribose) polymerase family, member 14/collaborator of STAT6 (Parp14, 1600029O10Rik, BC021340, KIAA1268, MGC29390, mKIAA1268) [BC021340] |
Unknown | [XM_207778] |
Histopathologic abnormalities and immunohistochemistry
Mouse | Meningitis | PV Cuffing | Inflammatory Lesions | Calcifications | Cysts | Inflammation With Cysts | |||||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Co | D | H | Ce | Co | D | H | Ce | Co | D | H | Ce | Co | D | H | Ce | Co | D | H | Ce | ||
U1 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 |
U2 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 |
U3 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 |
U4 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 |
U5 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 |
U6 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 |
I1 | 2 | 1 | 2 | 2 | 0 | 2 | 4 | 0 | 0 | 4 | 0 | 0 | 4 | 4 | 7 | 1 | 0 | 1 | 0 | 0 | 0 |
I2 | 1 | 1 | 1 | 2 | 2 | 2 | 0 | 1 | 0 | 0 | 0 | 0 | 4 | 2 | 0 | 0 | 0 | 0 | 0 | 0 | 0 |
I3 | 1 | 1 | 2 | 1 | 1 | 0 | 1 | 1 | 1 | 0 | 0 | 0 | 1 | 2 | 3 | 2 | 0 | 0 | 0 | 1 | 0 |
I4 | 2 | 1 | 3 | 2 | 2 | 0 | 1 | 1 | 1 | 0 | 0 | 0 | 0 | 3 | 5 | 1 | 0 | 0 | 1 | 1 | 0 |
I5 | 3 | 1 | 2 | 1 | 0 | 2 | 2 | 0 | 1 | 0 | 1 | 0 | 3 | 4 | 1 | 2 | 0 | 0 | 0 | 0 | 0 |
I6 | 3 | 1 | 1 | 3 | 1 | 1 | 1 | 4 | 0 | 0 | 2 | 0 | 3 | 2 | 2 | 2 | 1 | 0 | 0 | 1 | 0 |
I7 | 3 | 1 | 2 | 2 | 2 | 4 | 3 | 2 | 0 | 0 | 0 | 0 | 2 | 3 | 2 | 2 | 1 | 0 | 0 | 0 | 0 |
I8 | 1 | 1 | 1 | 2 | 2 | 1 | 0 | 1 | 0 | 0 | 0 | 0 | 1 | 0 | 1 | 1 | 0 | 0 | 0 | 0 | 0 |
Anatomic distribution of lesions and parasite burden demonstrate different areas of predominance of cysts and calcifications
P-values | Overall p-value* | |
---|---|---|
PV Cuffing (co) | 0.0003 | 0.0012 |
PV Cuffing (d) | 0.0010 | |
PV Cuffing (h) | 0.0009 | |
PV Cuffing (ce) | 0.0090 | |
Inflammatory lesions (co) | 0.0096 | 0.0012 |
Inflammatory lesions (d) | 0.0097 | |
Inflammatory lesions (h) | 0.0091 | |
Inflammatory lesions (ce) | 0.10 | |
Calcifications (co) | 0.39 | 0.0037 |
Calcifications (d) | 0.20 | |
Calcifications (h) | - | |
Calcifications (ce) | 0.0037 | |
Cysts (co) | 0.0035 | 0.0012 |
Cysts (d) | 0.0037 | |
Cysts (h) | 0.0032 | |
Cysts (ce) | 0.20 | |
Inflam w/cysts (co) | 0.39 | 0.051 |
Inflam w/cysts (d) | 0.39 | |
Inflam w/cysts (h) | 0.10 | |
Inflam w/cysts (ce) | - |
Co | D | H | Ce | p-value* | |
---|---|---|---|---|---|
PV Cuffing | 1 | 2 | 2 | 1.5 | 0.12 |
1-1 | 1–3 | 1–3 | 0–2 | ||
Inflammation | 1.5 | 1 | 1 | 0 | 0.37 |
0–4 | 0–4 | 0–4 | 0–1 | ||
Calcifications | 0 | 0 | 0 | 2.5 | 0.02 |
0–4 | 0–2 | 0-0 | 0–4 | ||
Cysts | 2.5 | 2 | 1.5 | 0 | < 0.01 |
0–4 | 0–7 | 0–2 | 0–1 | ||
Inflammation w/cysts | 0 | 0 | 0 | 0 | 0.70 |
0–1 | 0–1 | 0–1 | 0-0 | ||
Overall** | 4.5 | 6 | 4.5 | 4 | 0.61 |
2–12 | 1–13 | 3–10 | 3–6 |
Co | D | H | Ce | |
---|---|---|---|---|
Grooming | 0.52 | 0.62 |
0.78
| 0.54 |
Body Position |
0.76
|
0.92
|
0.87
| 0.66 |
Piloerection |
0.93
|
0.94
|
0.94
|
0.93
|
Tail Damage |
0.84
|
0.71
|
0.74
|
0.89
|
Tremor | 0.27 | 0.57 |
0.81
| 0.39 |
Palebral Closure | 0.17 | 0.49 | 0.56 | 0.14 |
Lacrimation | 0.42 | 0.57 | 0.57 | 0.30 |
Weight | -0.23 | -0.29 | -0.28 | -0.26 |
Spatial Locomotion | -0.29 | -0.41 | -0.69 | -0.45 |
Gait | 0.42 | 0.56 | 0.38 | 0.28 |
Urination |
0.93
|
0.94
|
0.94
|
0.93
|
Sniffs |
-0.78
|
-0.71
| -0.53 | -0.66 |
Rears |
-0.71
| -0.56 | -0.52 |
-0.72
|
Center | -0.69 | -0.45 | -0.41 |
-0.71
|
Freezing | 0.27 | 0.55 |
0.82
| 0.42 |
Latency to Move | 0.67 | 0.44 | 0.35 | 0.66 |
Grip Strength |
-0.78
| -0.57 | -0.49 |
-0.76
|
Pain Insensitivity | 0.64 | 0.65 |
0.72
| 0.67 |