Baricitinib for Reduction of HIV - CNS

This method is based on a proprietary protocol which is used in conjunction with a commercial HIV-1 RNA easyQ reagent kit (bioMerieux Inc, Lyon, France). Briefly, a specimen of up to 2 ml of human plasma or CSF is added to lysis buffer containing guanidine thiocyanate. HIV-1 RNA is extracted in combination with the easyMAG platform (bioMerieux, Inc). Eluates containing HIV-1 RNA are aliquoted into 0.5 mL reaction tubes and amplified using 3 enzymes: T7 RNA polymerase, avian myeloblastosis virus reverse transcriptase, and RNase H. Molecular beacons targeting the pol/gag region of HIV-1 RNA are used for amplification and detection by isothermal reactions at 41oC. HIV-1 RNA level is quantified in conjunction with the NucliSENS easyQ HIV-1 v2.0 Director software and a proprietary algorithm developed by bioMONTR Labs. The dynamic range of this HIV-1 assay is 1-5,000,000 copies/mL of 0.73 - 1600 HIV-1 copies /μl. The HIV-1 copy number is normalized per one million of WBCs.

The study team will obtain up to 40 milliliters (ml) of CSF from individuals who weigh at least 60 kilograms, otherwise we will draw up to 30 ml of CSF.

HIV Tat in the CSF will be quantified according to published methods for ELISA to measure HIV Tat in human CSF.

The study team will obtain up to 40 milliliters (ml) of CSF from individuals who weigh at least 60 kilograms, otherwise, they will draw up to 30 ml of CSF.

Briefly, after the isolation of Peripheral blood mononuclear cells (PBMC) and cells from CSF, samples will be resuspended in phosphate-buffered saline and stored for shipment in O-ring tubes. RNA will then be extracted using the Maxwell rapid sample concentrator (RSC) automated extraction platform. White Blood Cells (WBC) will be counted after red blood cell lysis using TC20 Automated Cell Counter, and these counts will be used to normalize the HIV-1 RNA results as copy numbers/106 cells. HIV-1 copy number per patient sample, in duplicate, is determined from a standard curve generated with HIV-1 plasmid controls, with a range of 0.73 - 1600 HIV-1 copies /μl. The HIV-1 copy number is normalized per one million of WBCs.

The study team will obtain up to 40 milliliters (ml) of CSF from individuals who weigh at least 60 kilograms, otherwise they will draw up to 30 ml of CSF.

Briefly, after the isolation of Peripheral blood mononuclear cells (PBMC) and cells from CSF, samples will be resuspended in phosphate-buffered saline and stored for shipment in O-ring tubes. DNA will then be extracted using the Maxwell rapid sample concentrator (RSC) automated extraction platform. WBCs will be counted after red blood cell lysis using TC20 Automated Cell Counter, and these counts will be used to normalize the HIV-1 DNA results as copy numbers/106 cells. HIV-1 copy number per patient sample, in duplicate, is determined from a standard curve generated with HIV-1 plasmid controls, with a range of 0.73 - 1600 HIV-1 copies /μl. The HIV-1 copy number is normalized per one million of WBCs.

The study team will obtain up to 40 milliliters (ml) of CSF from individuals who weigh at least 60 kilograms, otherwise we will draw up to 30 ml of CSF.

Cell associated DNA will be performed according to standard existing protocols using real-time PCR with a limit of detection of five copies.

Changes in the blood between baseline and week ten after study medication. Briefly, after isolation of Peripheral blood mononuclear cells (PBMC) and cells from patient's blood sample, samples will be resuspended in phosphate buffered saline and stored for shipment in O-ring tubes. RNA will then be extracted using the Maxwell rapid sample concentrator (RSC) automated extraction platform. WBCs will be counted after red blood cell lysis using TC20 Automated Cell Counter, and these counts will be used to normalize the HIV-1 RNA results as copy numbers/106 cells. HIV-1 copy number per patient sample, in duplicate, is determined from a standard curve generated with HIV-1 plasmid controls, with a range of 0.73 - 1600 HIV-1 copies /μl. The HIV-1 copy number is normalized per one million of WBCs.

Changes in the blood between baseline and week ten after study medication. Briefly, after isolation of Peripheral blood mononuclear cells (PBMC) and cells from the patient's blood sample, samples will be resuspended in phosphate buffered saline and stored for shipment in O-ring tubes. DNA will then be extracted using the Maxwell rapid sample concentrator (RSC) automated extraction platform. WBCs will be counted after red blood cell lysis using TC20 Automated Cell Counter, and these counts will be used to normalize the HIV-1 DNA results as copy numbers/106 cells. HIV-1 copy number per patient sample, in duplicate, is determined from a standard curve generated with HIV-1 plasmid controls, with a range of 0.73 - 1600 HIV-1 copies /μl. The HIV-1 copy number is normalized per one million of WBCs.

Change in the blood between baseline and week ten after study medication. Cell associated DNA will be performed according to standard existing protocols using real-time PCR with a limit of detection of five copies.

Change in the blood between baseline and week ten after study medication. The study team will obtain up to 40 milliliters (ml) of CSF from individuals who weigh at least 60 kilograms, otherwise we will draw up to 30 ml of CSF.

Blood integrated proviral DNA (IPDA and Alu-PCR) will be assessed: Integrated proviral DNA will be performed using existing methods in our group, which employ the Alu-PCR real-time platform, with a limit of 3 copies. Integrated proviral DNA assay (IPDA) to measure the replication competent reservoir will be used with digital droplet PCR methodology

This method is based on a proprietary protocol which is used in conjunction with a commercial HIV-1 RNA easyQ reagent kit (bioMerieux Inc, Lyon, France). Briefly, a specimen of up to 2 ml of human plasma or CSF is added to lysis buffer containing guanidine thiocyanate. HIV-1 RNA is extracted in combination with the easyMAG platform (bioMerieux, Inc). Eluates containing HIV-1 RNA are aliquoted into 0.5 mL reaction tubes and amplified using 3 enzymes: T7 RNA polymerase, avian myeloblastosis virus reverse transcriptase, and RNase H. Molecular beacons targeting the pol/gag region of HIV-1 RNA are used for amplification and detection by isothermal reactions at 41oC. HIV-1 RNA level is quantified in conjunction with the NucliSENS easyQ HIV-1 v2.0 Director software and a proprietary algorithm developed by bioMONTR Labs. The dynamic range of this HIV-1 assay is 1-5,000,000 copies/mLof 0.73

Change in HIV and inflammation levels in the blood between baseline and week ten after study medication.

Blood HIV antibodies by LIPS assay (gp120, integrase, protease, and p24) will be assayed. The LIPS assay uses recombinant chimeric fusion proteins with luciferase producing light and specific antigens for quantitative detection of antibodies.

Change in HIV and inflammation levels in the blood between baseline and week ten after study medication Fresh Human peripheral blood mononuclear cells (PBMC) will be cultured in the presence or absence of overlapping peptides which induces broad, non-specific T-cell activation. In addition to cytokine production (focusing on TNFα, IFNγ, and IL-6 measured with Quanterix technology). Researchers will measure cell surface markers by flow cytometry.

Change in levels in the blood between baseline and week ten after study medication.

The study team will measure CXCL10 from participants blood samples, in the setting of HIV using single molecule array technology by Quanterix.

Change in levels in the blood between baseline and week ten after study medication.

The study team will measure neopterin in the setting of HIV using the enzyme linked immunosorbent assays produced by Uman diagnostics, R\&D Systems (ultra-sensitive), and Themo Fisher.

Change in levels in the blood between baseline and week ten after study medication.

This will be measured using mesoscale discovery platform.

Change in levels in the blood between baseline and week ten after study medication.

This will be measured using mesoscale discovery platform.

In psychology, the Stroop effect is the delay in reaction time between congruent and incongruent stimuli.

The effect has been used to create a psychological test (the Stroop test) that is widely used in clinical practice and investigation.

A basic task that demonstrates this effect occurs when there is a mismatch between the name of a color (e.g., "blue", "green", or "red") and the color it is printed on (i.e., the word "red" printed in blue ink instead of red ink). When asked to name the color of the word it takes longer and is more prone to errors when the color of the ink does not match the name of the color.

Word, color, and color-word T-Scores of 40 or less are considered "low." Word, color, and color-word T-Scores above 40 or are considered "normal." In order for one score to be considered "higher" or "lower" than another, a 10 point or greater T score difference is required.

This will be performed in a quiet exam room. Participants will have the opportunity to take breaks as needed and snacks will be provided.

The Hopkins Verbal Learning Test-Revised (HVLT-R) is used to assess acquisition and delayed recall. The HVLT-R is a list learning test, which consists of 12 nouns within three semantic groups. The acquisition variable consists of three acquisition trials in which the administrator reads the words aloud and then asks the child to repeat as many as he/she can remember in any order. A delayed recall trial is introduced after 20-25 min, in which the child is asked to simply retrieve as many of the words listed in the acquisition trial as he/she can remember. The children are not informed about the delayed recall trial beforehand. Lower raw scores indicate difficulties with the task.

The Brief Visuospatial Memory Test (BVMT) is a commonly used assessment tool to measure visuospatial learning and memory abilities.

In three Learning Trials, the respondent views the stimulus page for 10 seconds and is asked to draw as many of the figures as possible in their correct location on a page in the response booklet. A Delayed Recall Trial is administered after a 25-minute delay. Last, a Recognition Trial, in which the respondent is asked to identify which of 12 figures were included among the original geometric figures, is administered.

An optional Copy Trial may be administered to screen for severe visuoconstructive deficits and to help in scoring recall responses.

12 scores may be derived from BVMT-R performance. Recall performance is recorded for each of the immediate recall trials (Trial 1, Trail 2, and Trial 3) and for the delayed recall trial (Delayed Recall).

The recall scores are combined to form three additional summary measures of learning and memory.

The Trail Making Test is a neuropsychological test of visual attention and task switching. It requires a subject to connect a sequence of 25 consecutive targets on a sheet of paper or computer screen. There are 2 parts to the test: in the first, the targets are all numbers from 1 to 25 and the test taker needs to connect them in sequential order; in the second part, the dots go from 1 to 13 and include letters from A to L. As in the first part, the patient must connect the dots in order while alternating letters and numbers, as in 1-A-2-B-3-C..., in the shortest time possible without lifting the pen from the paper. If the subject makes an error, the administrator corrects them before moving on to the next dot.

The subject's goal is to finish both parts as quickly as possible, with the time taken to complete the test being used as the primary performance metric. Scoring is based on time taken to complete the test (e.g. 35 seconds yielding a score of 35) with lower scores being better

This will be performed in a quiet exam room. Participants will have the opportunity to take breaks as needed and snacks will be provided.

The Digit Symbol Substitution Test (DSST) is a paper-and-pencil cognitive test presented on a single sheet of paper that requires a subject to match symbols to numbers according to a key located on the top of the page. The subject copies the symbol into spaces below a row of numbers. The number of correct symbols within the allowed time, usually 90 to 120 seconds, constitutes the score.

This will be performed in a quiet exam room. Participants will have the opportunity to take breaks as needed and snacks will be provided.

Letter-number sequencing is a subtest of the WAIS (Wechsler Adult Intelligence Scale) and the WISC (Wechsler Intelligence Scale for Children). It is a test that measures an individual's short-term memory skills in being able to process and re-sequence information. The task involves hearing a series of letters and digits, and then reporting back the stimuli with the letters in alphabetical order, and digits in ascending numerical order. The task is traditionally administered orally, but recent studies have administered versions of the tasks visually by displaying stimuli on a computer screen. Results suggest that performance differences on the Letter-Number Sequencing task may arise as a function of language background and task administration modality

This will be performed in a quiet exam room. Participants will have the opportunity to take breaks as needed and snacks will be provided.

This will be performed in a quiet exam room. Participants will have the opportunity to take breaks as needed and snacks will be provided.

The Grooved Pegboard Test (GPT) is used primarily as a measure of motor functioning, but some research indicates that performance on this test my also reflect cognitive factors, particularly attention and executive functioning.

This manipulative dexterity test contains twenty-five holes with randomly positioned slots and pegs which have a key along one side. Pegs must be rotated to match the hole before they can be inserted.

This procedure measures performance speed in a fine motor task by examining both sides of the body, inferences may be drawn regarding possible lateral brain damage.

The test requires more complete visual motor coordination than most of our pegboards and has been used in several neuropsychological test batteries, in student labs, and as a screening technique in industrial environment.

This will be performed in a quiet exam room. Participants will have the opportunity to take breaks as needed and snacks will be provided.

This will be performed in a quiet exam room. Participants will have the opportunity to take breaks as needed and snacks will be provided. The BDI-II is a widely used 21-item self-report inventory measuring the severity of depression in adolescents and adults.

The questionnaire consists of 21 questions with answers ranging from 0 to 3. Scores are later analyzed by adding all scores and comparing to ranges :

0-10:These ups and downs are considered normal 11-16:Mild mood disturbance 17-20:Borderline clinical depression 21-30:Moderate depression 31-40: Severe depression over 4: Extreme depression

Brain structure and function will be evaluated using magnetic resonance (MR) data acquired on a 7T Siemens Terra scanner with a multi-channel head coil. Data acquisition will include sequences for structural imaging, resting state functional MRI, diffusion imaging, perfusion imaging, and magnetic resonance spectroscopy.