Lyme bacteria survive 28-day course of antibiotics months after infection

https://www.sciencedaily.com/releases/2017/12/171213143613.htm

December 13, 2017

Bay Area Lyme Foundation, a leading sponsor of Lyme disease research in the US, today announced results of two papers published in the peer-reviewed journals PLOS ONE and American Journal of Pathology, that seem to support claims of lingering symptoms reported by many patients who have already received antibiotic treatment for the disease.

Based on a single, extensive study of Lyme disease designed by Tulane University researchers, the study employed multiple methods to evaluate the presence of Borrelia burgdorferi spirochetes, the bacteria that cause Lyme disease, before and after antibiotic treatment in primates. The study also measured the antibody immune response to the bacteria both pre- and post- treatment, as this is how current diagnostics typically evaluate Lyme disease in humans.

The data show that living B. burgdorferi spirochetes were found in ticks that fed upon the primates and in multiple organs after treatment with 28 days of oral doxycycline. The results also indicated that the immune response to the bacteria varied widely in both treated and untreated subjects.

"It is apparent from these data that B. burgdorferi bacteria, which have had time to adapt to their host, have the ability to escape immune recognition,tolerate the antibiotic doxycycline and invade vital organs such as the brain and heart," said lead author Monica Embers, PhD, assistant professor of microbiology and immunology at Tulane University School of Medicine.

"In this study, we were able to observe the existence of microscopic disease and low numbers of bacteria, which would be difficult to 'see' in humans but could possibly be the cause of the variable and nonspecific symptoms that are characteristic of post-treatment Lyme disease syndrome. Although current antibiotic regimens may cure most patients who are treated early, if the infection is allowed to progress, the 28-day treatment may be insufficient, based on these findings," Embers said.

The findings also demonstrated:

*  All subjects treated with antibiotics were found to have some level of infection 7 -- 12 months post treatment.
*  Despite testing negative by antibody tests for Lyme disease, two of 10 subjects were still infected with Lyme bacteria in heart and bladder.
*  Lyme bacteria which persist are still viable.

To better elucidate previous animal studies demonstrating that some B. burgdorferi bacteria survive antibiotics, the study explored Lyme disease infection in rhesus macaque primates treated with antibiotics and a control group who were also infected but not treated. This species has been shown to demonstrate a progression of Lyme disease most similar to humans, particularly related to erythema migrans, carditis, arthritis, and neuropathy of the peripheral and central nervous systems.

"Clearly, some medical practices governing diagnosis and treatment of Lyme disease should be reconsidered in light of this study. This study shows that we must reevaluate the current paradigm of antibody response tests for diagnosis and move away from the one size fits all approach to Lyme treatment," said Wendy Adams, Research Grant Director, Bay Area Lyme Foundation. "Every day, patients with Lyme disease are told their symptoms cannot be caused by Lyme, because they test negative on antibody tests or because they have received a single course of antibiotics. More research and funding are imperative."

In the study, ticks carrying B. burgdorferi spirochetes fed on ten primates. Four months post infection, half of the primates (five) received the antibiotic doxycycline orally for 28 days at a proportional dose to that used in human treatment. Five subjects were treated with placebo and all ten were evaluated by more than five different diagnostic methods to characterize any remaining infection. The researchers used several important techniques, including xenodiagnoses, to determine if the spirochete bacteria persisted.

The results show:

*  Few subjects displayed a rash. Although all subjects were infected, only one of the 10 displayed a rash with central clearing, the classical "bulls-eye" rash. The subject that developed this rash, interestingly, never mounted an immune response to five borrelia antigens throughout the study period, prior to and following treatment.
*  Organs may be infected even if antibody tests are negative. One subject which tested negative for B. burgdorferi by skin biopsy cultures, PCR and in vivo cultures, was found to have B. burgdorferi infecting the heart. Another untreated subject, who was ultimately shown to have residual Lyme bacteria in the bladder, showed a decrease in immune response over the course of infection, with a negative xenodiagnosis test in the late stage, which would signal that the animal self-cured.
*  Intact spirochetes were found in three of five treated and four of five untreated subjects based on xenodiagnosis results 12 months after the tick bite.
*  Immune responses to B. burgdorferi varied greatly post-treatment, with one subject's antibody levels dropping to pre-bite levels for three antigens while another subject experienced elevated antibodies for the same antigens throughout the study period. This is significant because it demonstrates that subjects infected with the same strain of B. burgdorferi may have different immune responses to the same antigen. And, because humans, like primates, are genetically diverse, it underscores that testing antibody responses may be inherently unreliable as a singular diagnostic modality for Lyme disease.
* Widespread and variable microscopic disease was observed in all infected subjects, despite antibiotic treatment. Compared to uninfected subjects of the same age, infected subjects in this study (treated and untreated) demonstrated Inflammation in and around the heart, in skeletal muscles, joints, and the protective sheath that covers the brain, and near peripheral nerves.
*   Rare, but intact B. burgdorferi spirochetes were found in the tissues of both the treated and untreated subjects. In two subjects treated with doxycycline, multiple Lyme bacteria were observed in the brain tissue. Others organs in which the spirochetes were observed included the heart, joints, bladder, skeletal muscle and adjacent to peripheral nerves.

*******
Ehkä vähitellen järki voittaa. Nykyisin jos potilaalla oireet jatkuvat, se on potilaan syy, sen päässä on vikaa, koska borrelioosi paranee kahden viikon doxy kuurilla..

Epstein-Barr virus is present in the brain of most cases of multiple sclerosis and may engage more than just B cells

https://www.ncbi.nlm.nih.gov/pubmed/29394264#

PLoS One. 2018 Feb 2

Abstract

Multiple sclerosis (MS) is a chronic neuroinflammatory condition of the central nervous system (CNS). It is a major cause of neurological disability in young adults, particularly women. What triggers the destruction of myelin sheaths covering nerve fibres is unknown. Both genetic and infectious agents have been implicated. Of the infectious agents, Epstein-Barr virus (EBV), a common herpesvirus, has the strongest epidemiological and serological evidence. However, the presence of EBV in the CNS and demonstration of the underlying mechanism(s) linking EBV to the pathogenesis of MS remain to be elucidated. We aimed at understanding the contribution of EBV infection in the pathology of MS. We examined 1055 specimens (440 DNA samples and 615 brain tissues) from 101 MS and 21 non-MS cases for the presence of EBV using PCR and EBER-in situ hybridization (EBER-ISH). EBV was detected by PCR and/or EBER-ISH in 91/101 (90%) of MS cases compared to only 5/21 (24%) of non-MS cases with other neuropathologies. None of the samples were PCR positive for other common herpesviruses (HSV-1, CMV, HHV-6). By quantitative PCR, EBV viral load in MS brain was mainly low to moderate in most cases. However, in 18/101 (18%) of MS cases, widespread but scattered presence of EBV infected cells was noted in the affected tissues by EBER-ISH. Immunohistochemical analysis of EBV gene expression in the 18 heavily infected cases, revealed that the EBV latent protein EBNA1, and to a lesser extent the early lytic protein BZLF1 were expressed. Furthermore, using double-staining we show for the first time that astrocytes and microglia, in addition to B-cells can also be infected. To the best of our knowledge, this is the most comprehensive study demonstrating that EBV is present and transcriptionally active in the brain of most cases of MS and supports a role for the virus in MS pathogenesis. Further studies are required to address the mechanism of EBV involvement in MS pathology.

New clues to Epstein-Barr virus

https://www.sciencedaily.com/releases/2013/02/130221152738.htm

February 21, 2013
Epstein-Barr virus (EBV) affects more than 90 percent of the population worldwide and was the first human virus found to be associated with cancer. Now, researchers from Beth Israel Deaconess Medical Center (BIDMC) have broadened the understanding of this widespread infection with their discovery of a second B-cell attachment receptor for EBV.

The new findings, which currently appear on-line in Cell Reports, reinforce current directions being taken in the development of a vaccine to guard against EBV, and raise important new questions regarding the virus's possible relationship to malaria and to autoimmune diseases.

"Our discovery that CD35 is an attachment receptor for EBV helps explain several previously unsolved observations," explains the study's senior author Joyce Fingeroth, MD, a member of the Division of Infectious Disease at BIDMC and Associate Professor of Medicine at Harvard Medical School.

First discovered in the early 1960s, EBV is one of eight viruses in the human herpesvirus family. The virus affects nine out of 10 people at some point in their lifetimes.

Infections in early childhood often cause no disease symptoms, but people infected during adolescence or young adulthood may develop infectious mononucleosis. EBV is also associated with several types of cancer, including Hodgkin's lymphoma, non-Hodgkin's lymphoma and nasopharyngeal carcinoma, and has been linked to certain autoimmune disorders.

"EBV was the first human virus that was discovered to be a tumor virus," explains Fingeroth. "In fact, individuals who have had infectious mononucleosis have a four times increased risk of developing Hodgkin's disease." After the initial infection, the EBV virus remains in a person's body for life.

To gain entry, viruses must first attach to their host cells. For herpesviruses, receptors on the viral envelope become connected to complementary receptors on the cell membrane. In the case of EBV, the virus gains access to the immune system by attaching to primary B cells.

Nearly 30 years ago, Fingeroth and her colleagues discovered that this attachment occurs via the CD21 protein, which until now was the only known B cell attachment receptor for EBV. The recent finding that B cells from a patient lacking CD21 can be infected and immortalized by EBV had indicated that an alternative attachment receptor must exist. The identification of this second receptor -- CD35 -- by Fingeroth's team, led by first author Javier Ogembo, PhD, of BIDMC and the University of Massachusetts Medical School, not only underscores an important finding regarding primary infection but also underscores the importance of EBVgp350/220, (the virus protein that has been found to bind to both attachment receptors) for the development of a vaccine against EBV.

"The EBV glycoprotein gp350/220, is the most abundant surface glycoprotein on the virus," says Fingeroth, adding that these results further suggest the virus fusion apparatus is the same for both receptors. "An EBV vaccine might be able to prevent infection or, alternatively, greatly reduce a person's risk of developing infectious mononucleosis and EBV-associated cancers, without necessarily preventing the EBV infection itself," she explains.

Interestingly, whereas a human has now been identified to be lacking the CD21 receptor, no persons are known to lack CD35. "CD35 is a latecomer in evolution and in its current form, exists only in humans," says Fingeroth. "We know that it is often targeted in autoimmune diseases and was recently identified as a malaria receptor. Our new discovery may, therefore, reveal new avenues for the exploration of unexplained links between EBV, autoimmune disease, malaria, and cancer."

In addition to Fingeroth and Ogembo, study coauthors include BIDMC investigators Lakshmi Kannan, Ionita Ghiran, Anne Nicholson-Weller and George Tsokos; and UMass investigator Robert Finberg.

Experimental immunotherapy ATX-MS-1467 shows promising trial results

https://msra.org.au/news/experimental-immunotherapy-atx-ms-1467-shows-promising-...
trial-results/

08 March, 2018

   Unlike most available MS medications which suppress or modify the immune system, immunotherapies aim to ‘re-educate’ the cells of the immune system to prevent them responding to a specific target.
   Experimental immunotherapy ATX-MS-1467 aims to re-train the immune system’s response to myelin, and previously showed good results in laboratory models of MS
   A Phase 1b clinical trial tested the safety of ATX-MS-1467 and found it to be well tolerated
   A Phase 2a clinical trial showed that the treatment was able to reduce lesions on magnetic resonance imaging (MRI)

MS results from the immune system mistakenly attacking and destroying myelin, the protective layer that insulates nerves in the brain and spinal cord. Most current treatments available for MS act by suppressing or ‘resetting’ the immune system to stop this attack. Immunotherapy is a completely different approach to treating MS. Immunotherapy aims to ‘re-educate’ the cells of the immune system responsible for the MS attack to reduce the likelihood of a relapse.

Results of two clinical trials of an experimental new immunotherapy for people with MS have recently been published in the journal Neurology showing encouraging results.

Myelin is made up of a range of fat molecules and some proteins. In MS, the immune cells mistakenly target some of the protein components of the myelin, including parts of one protein known as myelin basic protein. The experimental immunotherapy, ATX-MS-1467, is made up of a cocktail of four different protein fragments from myelin basic protein. The protein fragments are injected under the skin and are able to ‘teach’ immune cells not to attack myelin basic protein.

This essentially re-trains the immune system to recognise myelin as ‘self’ with the hope of preventing the myelin damage seen in MS. Previous research had shown good results of ATX-MS-1467 in a laboratory model of MS. In the studies just published, the researchers conducted clinical trials in a small number of people with relapsing MS to test for safety (Phase 1b trial) and effectiveness (Phase 2a trial).

The Phase 1b clinical trial compared two routes of administration of the drug, intradermal injections (injections into the skin just below the outermost layer) and subcutaneous injections (deeper injections into the fat layer below the skin) and compared a range of doses in 43 people with relapsing MS. Participants were given an injection every two weeks for 16 weeks and then followed for a further 32 weeks. This study showed that while there were some mild and moderate side effects, ATX-MS-1467 was well tolerated and that the intradermal injections were a better way of delivering the immunotherapy.

Exploratory investigations of the effectiveness of the treatment in the Phase I study indicated that it may have some temporary effect in reducing the gadolinium enhancing lesions on magnetic resonance imaging (MRI), a marker of active disease but lesions returned to pre-treatment levels after the 32 weeks off the medication.

The Phase 2b clinical trial was then designed to more thoroughly examine the effectiveness of the treatment in 37 participants compared with baseline levels of disease. Participants were given treatment over 16 weeks and then followed-up for 16 weeks off treatment. As for the first trial, the Phase 2b trial showed that ATX-MS-1467 was able to reduce gadolinium enhancing lesions on MRI at the end of the treatment period but also showed that this reduction persisted over the shorter 16 weeks of follow-up. The Phase 2b trial also showed that ATX-MS-1467 reduced the number of new or enlarging lesions on MRI.

Participants did not have significant reductions in their expanded disability status scale (EDSS) or MS functional composite (MSFC) measurements, both of which assess levels of disability in the clinic, as a result of the treatment. However, the short duration of the trial would make it difficult to reliably detect any changes in disability.

These promising results imply re-training the immune system to prevent it’s attacks on myelin may be an effective strategy and further clinical trial testing of ATX-MS-1467 is warranted in larger groups of people with MS.

HLA alleles modulate EBV viral load in multiple sclerosis.

https://www.ncbi.nlm.nih.gov/pubmed/29587799

J Transl Med. 2018 Mar 27;

Abstract
BACKGROUND:

The etiopathology of multiple sclerosis (MS) is believed to include genetic and environmental factors. Human leukocyte antigen (HLA) alleles, in particular,  are associated with disease susceptibility, whereas Epstein Barr Virus (EBV) infection has long been suspected to play a role in disease pathogenesis. The aim of the present study is to evaluate correlations between HLA alleles and EBV infection in MS.

METHODS:

HLA alleles, EBV viral load (VL) and serum anti-EBV antibody titers were evaluated in EBV-seropositive MS patients (N = 117) and age- and sex-matched healthy controls (HC; N = 89).

RESULTS:

Significantly higher DNA viral loads (p = 0.048) and EBNA-1 antibody titer (p = 0.0004) were seen in MS compared to HC. EBV VL was higher in HLA-B*07+ (p = 0.02) and HLA-DRB1*15+ (p = 0.02) MS patients, whereas it was lower in HLA-A*02+ (p = 0.04) subjects. EBV VL was highest in HLA-A*02-/B*07+/DRB1*15+ patients and lowest in HLA-A*A02+/B*07-/DRB1*15- individuals (p < 0.0001). HLA-B*07 resulted the most associated allele to EBV VL after multiple regression analysis considering altogether the three alleles, (p = 0.0001). No differences were observed in anti-EBV antibody titers in relationship with HLA distribution.

CONCLUSIONS:

Host HLA-B*07 allele influence EBV VL in MS. As HLA-class I molecules present antigens to T lymphocytes and initiate immune response against viruses, these results could support a role for EBV in MS.

******
Eli, jos ymmärränn oikein, MS:ään liitetyt geenit ovat syyllisiä heikkoon immuunivastukseen EBV-virukselle.

Different diseases elicit distinct sets of exhausted T cells

https://www.sciencedaily.com/releases/2018/05/180515113754.htm

May 15, 2018
The battle between the human immune system and long-term, persisting infections and other chronic diseases such as cancer results in a prolonged stalemate. Over time battle-weary T cells become exhausted, giving germs or tumors an edge. Using data from multiple molecular databases, researchers from the Perelman School of Medicine at the University of Pennsylvania have found nine distinct types of exhausted T cells ("Tex"), which could have implications for fighting chronic infections, autoimmunity, and cancer. They published their findings in Immunity this week.

"Exhausted T cells are a discrete cell lineage that have become important immunotherapy targets for chronic infection and cancer," said senior author John Wherry, PhD, a professor of Microbiology and director of the Institute for Immunology. "Now, we know that exhausted T cells are a vastly diverse set of immune cells."

Wherry's lab has spent the last decade describing these populations of fatigued cells. Overall, when normal T cells become exhausted, they develop defects in their germ- and tumor-fighting capabilities. Tex also express inhibitory receptor proteins on their surface that stall key biochemical pathways, provoke changes in control of gene expression, alter metabolism for making energy to fight infections and tumors, and prevent development of optimal immune function.

New, highly effective immunotherapies that target these inhibitory receptors expressed by Tex such as PD-1 or CTLA-4 have shown dramatic effects among patients with melanoma and other diseases, with potential to also combat breast, ovarian and other cancers. Although Tex have been implicated in the response to checkpoint blockade drugs in animal models, the underlying immunological mechanisms of their therapeutic response or failure in people is only now being studied in earnest.

"Exhausted T cells are quite diverse, as are all types of T cells," Wherry said. "This sheer diversity is the hallmark of the human immune system that has to essentially have a way to respond to every germ an individual might encounter in a lifetime."

Knowing this, the Penn team asked what the diversity in the Tex pool reveals about a disease itself and its course in a patient. They developed an assay to investigate the molecules that control gene expression in Tex by comparing them to other types of T cells and within a Tex population in blood from HIV patients whose viral load is well-controlled.

Next, they defined core exhaustion-specific genes and identified disease-induced molecular changes in Tex populations in HIV with uncontrolled disease and in human lung cancer. Using this data, the Tex fell into nine distinct clusters of similar expression patterns with regard to transcription factors and inhibitory receptors.

Because of the clusters' relationships to specific disease type and progression, the team's aim is to use the signature of a Tex cluster to assess a patient's overall immune health and likelihood of responding to a certain therapy. "We want to be able to select and tailor immune therapies according to a patient's exhausted T cell pool and its individual characteristics," Wherry said.

Applying this type of assessment to exhausted T cells in the context of immunotherapy clinical trials might identify patients more likely to benefit from specific types of combination immunotherapies and may point to underlying mechanisms in the specific types of exhausted T cells responding to an infection or cancer.

Epstein Barr Virus found in lesions in the brains of people with MS

https://msra.org.au/news/epstein-barr-virus-found-lesions-brains-people-ms/

02 July, 2018

   Epstein Barr Virus (EBV) is the virus that causes glandular fever and is one of the known risk factors for developing MS.
   EBV infects B cells in the blood and new research has investigated EBV in the brains of people with MS.
   They identified that a higher proportion of MS brains showed evidence of EBV infected cells and that the cells moved from the bloodstream to the chronic lesions in the MS brain. Further strengthening the evidence for a role of EBV in MS.

Being infected by Epstein Barr Virus (EBV), the virus that causes glandular fever, is a known risk factor of developing MS. While the rate of EBV infection in the general population is very high, essentially everyone  with MS has had EBV at some point in their lives whether they realise or not. EBV infects immune cells called B cells in the blood, but whether EBV is found in the immune  cells that have entered in the brain during an MS attack is still not clear, as previous research was contradictory.

Now a new research paper, which includes some researchers from the Queensland Institute of Medical Research, has attempted to answer this question again and mapped EBV throughout the MS brain and in different types of MS lesions.

The study looked at brain tissue under the microscope from people with MS and compared it to brain tissue from people without MS. The people with MS had been diagnosed with different forms of the disease which meant the researchers could examine different types of lesions – both chronic lesions and chronic active lesions – in the brain tissue.

After infection, EBV can remain in the body in an inactive phase, which is known as ‘latent’ infection. A much higher proportion of the brains from people with MS contained a protein related to a latent EBV infection than the brains of those which did not have MS. In the brains of people with MS, the researchers could also see that the immune cells that respond to EBV infection had moved out of the blood and into the brain far more frequently. In the MS brains, these cells could be seen within the chronic MS lesions.

Another method was used to detect evidence of latent EBV infection within the brain, which tracks gene activity of the virus in a latent infection. This method showed that 6 of the 7 MS brains tested showed populations of these cells in different locations in the brain, but in brains without MS, they could only identify a single cell in 2 of the 4 brains tested.

This work has shown that B cells infected with EBV do enter travel to the brain and leave the blood circulatory system and enter the brain tissue. This happens in people with or without MS, but much more often in people who develop MS and this may suggest that people who go on to develop MS have a reduced capacity to deal with EBV infection. This idea that people with MS respond differently to EBV infection is not new and the immune response to EBV has been the subject of much research in the field of MS, particular by Professor Michael Pender and Professor Rajiv Khanna (an author on the study) from the University of Queensland. This research has led to a new approach to treatment of MS known as adoptive immunotherapy, which aims to boost person with MS’s immune response to infection with EBV using their own cells. This treatment is now in clinical trials for people with progressive MS and showing promising results . More recently, further trials were announced  that will test a similar treatment approach where cells are taken from donor rather than the person with MS.

Scientists find a new way to attack herpesviruses

https://www.sciencedaily.com/releases/2018/08/180828121414.htm

August 28, 2018
Human cytomegalovirus is a leading cause of birth defects and transplant failures. As it's evolved over time, this virus from the herpes family has found a way to bypass the body's defense mechanisms that usually guards against viral infections. Until now, scientists couldn't understand how it manages to do so.

A team of scientists led by Leor S. Weinberger, PhD, the William and Ute Bowes Distinguished Professor and director of the Gladstone-UCSF Center for Cell Circuitry, uncovered the mechanism that allows the virus to replicate. Their study, published in the scientific journal PNAS, could open new therapeutic avenues to treat not only cytomegalovirus, but other viruses as well.

Normally, when a virus enters your cell, that cell blocks the virus's DNA and prevents it from performing any actions. The virus must overcome this barrier to effectively multiply.

To get around this obstacle, cytomegalovirus doesn't simply inject its own DNA into a human cell. Instead, it carries its viral DNA into the cell along with proteins called PP71. After entering the cell, it releases these PP71 proteins, which enables the viral DNA to replicate and the infection to spread.

"The way the virus operates is pretty cool, but it also presents a problem we couldn't solve," said Noam Vardi, PhD, postdoctoral scholar in Weinberger's laboratory and first author of the new study. "The PP71 proteins are needed for the virus to replicate. But they actually die after a few hours, while it takes days to create new virus. So how can the virus successfully multiply even after these proteins are gone?"

The researchers found that, while PP71 is still present in the cell, it activates another protein known as IE1. This happens within the first few hours of the virus entering the cell, allowing the IE1 protein to take over after PP71 dies and continue creating new virus.

To confirm their findings, the team created a synthetic version of the virus that allowed them to adjust the levels of the IE1 proteins using small molecules. With this technique, they could let the virus infect the cell while controlling how quickly the IE1 protein would break down in the cell.

"We noticed that when the IE1 protein degrades slowly, as it normally does, the virus can replicate very efficiently," said Vardi. "But if the protein breaks down faster, the virus can't multiply as well. So, we confirmed that the virus needs the IE1 protein to successfully replicate."

This study could have broad implications for the scientific community, which has been struggling to determine how cells maintain their identity over time. During development, for instance, stem cells choose a path based on the proteins that surround them. But even after these initial proteins disappear, the specialized cells don't change. So, stem cells that turn into neurons during development continue to be neurons long after those proteins are gone.

"The issue is similar for the virus," explained Weinberger, who is also a professor of pharmaceutical chemistry at UC San Francisco. "It was not clear what mechanisms allowed the virus to continue replicating long after the initial signal from the PP71 had decayed to a whisper. Our findings uncover a circuit encoded by the virus that controls its fate and indicate that such circuits may be quite common in viruses."

The new study could lead to a new therapeutic target to attack cytomegalovirus and other herpesviruses, such as Epstein-Barr virus that causes mononucleosis and herpes simplex virus 1 and 2 that produce most cold sores and genital herpes.