Research ArticleHIV

Heterogeneous antiretroviral drug distribution and HIV/SHIV detection in the gut of three species

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Science Translational Medicine  03 Jul 2019:
Vol. 11, Issue 499, eaap8758
DOI: 10.1126/scitranslmed.aap8758
  • Fig. 1 Antiretroviral drug distribution in gut tissue from three species.

    Representative images of antiretroviral drug distribution in a single cross-sectional 10-μm slice of ileum from an uninfected macaque, an HIV-infected humanized BLT mouse, and an HIV-infected human measured using MSI are shown. A total of 24 macaques, 98 mice, and 10 human tissue slices were imaged by MSI. (A to J) Macaque. (K to N) Mouse. (O to V) Human. Relative drug abundance is indicated by the color scale: Lighter colors (yellow and white) are associated with higher drug concentrations than darker colors (red and black). Endogenous cholesterol distribution is shown in macaque ileum in (A) to demonstrate tissue orientation for the remaining images. Drug distribution images for all antiretroviral drugs administered to this macaque (B to E) are overlaid with the localization of CD3+ T cells from an adjacent tissue section (F), and the colocalization is shown in (G) to (J) (drug, red; CD3+ T cells, green). Similar images of antiretroviral drug (tenofovir) concentration on its own or overlaid with CD3+ T cell localization are shown for an HIV-infected BLT mouse (K to N) and an HIV-infected human (O to V). Scale bars to the left of each endogenous cholesterol image apply to all antiretroviral drug distribution images for that tissue slice. In the BLT mouse ileum slice (K and L), tenofovir was the only drug detected by MSI and was overlaid with the CD3+ T cell image (N). In the human ileum tissue section (O to R), all three drugs contained in the patient’s antiretroviral drug regimen were detected by MSI. (T) to (V) show these images overlaid with CD3+ T cell localization (S).

  • Fig. 2 Incomplete antiretroviral drug and CD3+ T cell colocalization in macaque ileum.

    Representative images of antiretroviral drug distribution in a single cross-sectional 10-μm slice of ileum from an SHIV-infected macaque measured by MSI are shown. A total of 12 macaque ileal tissue slices were imaged by MSI. (B to D) Lighter colors (yellow and white) are associated with higher antiretroviral drug concentrations than darker colors (red and black). Endogenous cholesterol distribution is shown in (A) to demonstrate tissue orientation for the remaining images. (E) Composite image of the distribution of all three drugs. (G to J) These images were overlaid with the CD3+ T cell distribution from an adjacent ileal tissue slice (F) (drug, red; CD3+ T cells, green). The percentage of total CD3+ T cells colocalizing with tenofovir (G), maraviroc (H), or atazanavir (I) is shown at the bottom right of the respective panels. The percentage of total CD3+ T cells colocalizing with at least one drug is shown in (J).

  • Fig. 3 Antiretroviral drug concentrations and SHIV RNA expression.

    (B to E) Representative images of the distribution of four antiretroviral drugs in a single cross-sectional 10-μm slice of ileum from an SHIV-infected macaque measured by MSI. A total of 24 macaque tissue slices were imaged by MSI. Lighter colors (yellow and white) are associated with higher drug concentrations compared to darker colors (red and black). Endogenous cholesterol distribution is shown in (A) to enable tissue orientation for the remaining images. Images in (B) to (E) were overlaid (G to J) with a viral RNA image at matched spatial resolution from an adjacent slice (F) (drug, red; viral RNA, cyan). The percentage of total viral RNA colocalizing with drug is shown in the bottom left corner of each image. (K to N) Images show the distribution of each antiretroviral drug as a function of the drug’s IC90 value (concentrations above the IC90, green; concentrations below the IC90, red). (O to R) Similar analysis for drug that colocalizes with viral RNA.

  • Fig. 4 Colocalization of antiretroviral drugs with MDR1 efflux transporter expression in three species.

    Representative images of antiretroviral drug distribution in a single cross-sectional 10-μm slice of ileum from an uninfected macaque, an HIV-infected humanized BLT mouse, and an HIV-infected human are shown. A total of 24 macaque, 98 mouse, and 10 human tissue slices were imaged by MSI. Antiretroviral drug concentrations in (B) to (E) are indicated by the color scale on the left: Lighter colors (yellow and white) are associated with higher drug concentrations compared to darker colors (red and black). Endogenous cholesterol distribution is shown in (A) to demonstrate tissue orientation for the remaining images. Images (B) to (E) were overlaid with images of MDR1 efflux transporter expression in an adjacent tissue slice (F). (G to J) Overlaid images show drug in red and MDR1 transporter in green. Pearson correlation coefficients (r) between these variables are listed in the bottom left corner of each overlaid image. Images of drug distribution alone or overlaid with MDR1 expression are shown for a BLT HIV-infected mouse (J to M) and an HIV-infected human (N to U). Scale bars to the left of each endogenous cholesterol image apply to all antiretroviral drug distribution images from that tissue slice. In the BLT mouse ileum slice shown in (K) and (L), maraviroc was the only drug detected by MSI and is shown overlaid with MDR1 expression (N). In the human ileum tissue section shown in (O) to (R), all three drugs contained in the patient’s antiretroviral regimen were detected by MSI. Images of drug distribution are overlaid with MDR1 expression (S) in (T) to (V).

  • Table 1 Percent CD3+ T cells not colocalizing with any antiretroviral drug.

    Data are median and range.

    BLT (n = 13)hu-HSC-Rag (n = 36)Macaque (n = 12)Human (n = 5)P
    Ileum50 (10–100)100 (20–100)40 (10–90)40 (30–50)<0.01
    Rectum40 (10–100)100 (20–100)50 (20–80)40 (20–70)<0.01
  • Table 2 Percent viral RNA expression colocalizing with antiretroviral drug.

    Data are median and range.

    BLT (n = 7)hu-HSC-Rag
    (n = 18)
    Macaque
    (n = 6)
    Human
    (n = 5)*
    P
    Ileum10 (0–100)0 (0–100)80
    (40–90)
    90 (80–90)<0.01
    Rectum50 (20–100)0 (0–100)60
    (20–80)
    100
    (100–100)
    <0.01

    *Viral RNA was detected in rectal samples from only two of five women.

    • Table 3 Drug concentrations proximate to viral RNA.

      Data are median and range. Conversion from ng/ml to ng/g was performed using an assumed tissue density of 1.06 g/ml. IC90 values for each drug are reported from the following references: tenofovir (55), tenofovir diphosphate (56), emtricitabine (57), emtricitabine triphosphate (58), efavirenz (59), maraviroc (60), atazanavir (61), raltegravir (62). ND, analyte not detected in any tissue; TFV, tenofovir; TFVdp, tenofovir diphosphate; FTC, emtricitabine; FTCtp, emtricitabine triphosphate; RAL, raltegravir; EFV, efavirenz; MVC, maraviroc; ATZ, atazanavir.

      DrugIleumRectumReported
      in vitro
      IC90
      MiceAll macaquesAll
      humans
      MiceAll macaquesAll
      humans
      BLThu-HSC-RagBLThu-HSC-Rag
      TFV49 (0–1,782)0 (0–93)97 (0.4–151)0 (0–340)844 (0–1,499)0 (0–5,152)80 (0–182)0 (0–158)2,980
      TFVdp*1.7 (0–4.1)0.8 (0–7)0.4 (0.03–2.8)21 (11–65)0.003 (0–0.1)0.07 (0–11)0.2 (0.03–17)17 (9.4–23)1.7
      FTC0 (0–96)ND70 (1–206)ND0 (0–37)0 (0–41)0 (0–66)0 (0–6)84
      FTCtp*0.09 (0–0.2)ND0.02 (0.005–0.05)0.7 (0.1–6.5)NDND0.001 (0–0.2)3.3 (1.7–7.7)2.8
      RALNDND12,887 (4,420–21,353)18,632NDND6,891 (2,877–10,906)38710
      EFVND358 (215–501)24,6870 (0–21)385 (268–501)15,17810
      MVC5,739 (0–14,872)0 (0–4,776)3,199 (1,059–5,822)13,117 (1,589–8,759)0 (0–724)3,990 (957–4,625)ND6
      ATZ2,972 (0–28,616)0 (0–117,819)1,987 (1,152–3,674)ND596 (0–2,794)0 (0–23,632)1,068 (980–8,861)1,83211

      *Tenofovir diphosphate and emtricitabine triphosphate concentrations (detected by LC-MS) and IC90 values are reported as ×105 fmol/g of tissue.

      Supplementary Materials

      • stm.sciencemag.org/cgi/content/full/11/499/eaap8758/DC1

        Fig. S1. Effect of gelatin embedding on antiretroviral drug distribution.

        Fig. S2. Resolution matching of microscopy and MSI data.

        Fig. S3. Image colocalization workflow.

        Table S1. Subject demographics.

        Table S2. Human plasma and tissue antiretroviral drug concentrations.

        Table S3. Proportion of CD3+ T cells exposed to at least one antiretroviral drug.

        Table S4. Dosing of animals.

        Data file S1. Individual-level data for tables.

      • The PDF file includes:

        • Fig. S1. Effect of gelatin embedding on antiretroviral drug distribution.
        • Fig. S2. Resolution matching of microscopy and MSI data.
        • Fig. S3. Image colocalization workflow.
        • Table S1. Subject demographics.
        • Table S2. Human plasma and tissue antiretroviral drug concentrations.
        • Table S3. Proportion of CD3+ T cells exposed to at least one antiretroviral drug.
        • Table S4. Dosing of animals.

        [Download PDF]

        Other Supplementary Material for this manuscript includes the following:

        • Data file S1 (Microsoft Excel format). Individual-level data for tables.

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